TY  - DATA
AU  - Gligorijević, Nikola
AU  - Lujić, Tamara
AU  - Mutić, Tamara
AU  - Vasović, Tamara
AU  - Aćimović, Jelena
AU  - de Guzman, Maria Krishna
AU  - Stanić-Vučinić, Dragana
AU  - Ćirković-Veličković, Tanja
PY  - 2024
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6469
AB  - Research data for the unpublished paper: Ovalbumin interaction with polystyrene and polyethylene terephthalate microplastics alters its structural properties. Densitometry analysis of ovalbumin band intensity after digestion in the presence of microplastics on SDS-PAGE gels.
T1  - Research data no. 2 for the manuscript: Ovalbumin interaction with polystyrene and polyethylene terephthalate microplastics alters its structural properties
UR  - https://hdl.handle.net/21.15107/rcub_cherry_6469
ER  - 

@misc{
author = "Gligorijević, Nikola and Lujić, Tamara and Mutić, Tamara and Vasović, Tamara and Aćimović, Jelena and de Guzman, Maria Krishna and Stanić-Vučinić, Dragana and Ćirković-Veličković, Tanja",
year = "2024",
abstract = "Research data for the unpublished paper: Ovalbumin interaction with polystyrene and polyethylene terephthalate microplastics alters its structural properties. Densitometry analysis of ovalbumin band intensity after digestion in the presence of microplastics on SDS-PAGE gels.",
title = "Research data no. 2 for the manuscript: Ovalbumin interaction with polystyrene and polyethylene terephthalate microplastics alters its structural properties",
url = "https://hdl.handle.net/21.15107/rcub_cherry_6469"
}

Gligorijević, N., Lujić, T., Mutić, T., Vasović, T., Aćimović, J., de Guzman, M. K., Stanić-Vučinić, D.,& Ćirković-Veličković, T.. (2024). Research data no. 2 for the manuscript: Ovalbumin interaction with polystyrene and polyethylene terephthalate microplastics alters its structural properties. .
https://hdl.handle.net/21.15107/rcub_cherry_6469

Gligorijević N, Lujić T, Mutić T, Vasović T, Aćimović J, de Guzman MK, Stanić-Vučinić D, Ćirković-Veličković T. Research data no. 2 for the manuscript: Ovalbumin interaction with polystyrene and polyethylene terephthalate microplastics alters its structural properties. 2024;.
https://hdl.handle.net/21.15107/rcub_cherry_6469 .

Gligorijević, Nikola, Lujić, Tamara, Mutić, Tamara, Vasović, Tamara, Aćimović, Jelena, de Guzman, Maria Krishna, Stanić-Vučinić, Dragana, Ćirković-Veličković, Tanja, "Research data no. 2 for the manuscript: Ovalbumin interaction with polystyrene and polyethylene terephthalate microplastics alters its structural properties" (2024),
https://hdl.handle.net/21.15107/rcub_cherry_6469 .

TY  - DATA
AU  - Gligorijević, Nikola
AU  - Lujić, Tamara
AU  - Mutić, Tamara
AU  - Vasović, Tamara
AU  - de Guzman, Maria Krishna
AU  - Aćimović, Jelena
AU  - Stanić-Vučinić, Dragana
AU  - Ćirković-Veličković, Tanja
PY  - 2024
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6465
AB  - Data used for the analysis of pepsin interaction with polystyrene microplastic of 10 and 100 µm in size. Binding isotherms data were used for the construction of binding isotherms from which binding between pepsin and polystyrene microplastic was described. Structural features of pepsin in the presence of polystyrene were analysed by CD spectrometry and spectrofluorimetry.
T1  - Research data no. 1 for the manuscript: Ovalbumin interaction with polystyrene and polyethylene terephthalate microplastics alters its structural properties
UR  - https://hdl.handle.net/21.15107/rcub_cherry_6465
ER  - 

@misc{
author = "Gligorijević, Nikola and Lujić, Tamara and Mutić, Tamara and Vasović, Tamara and de Guzman, Maria Krishna and Aćimović, Jelena and Stanić-Vučinić, Dragana and Ćirković-Veličković, Tanja",
year = "2024",
abstract = "Data used for the analysis of pepsin interaction with polystyrene microplastic of 10 and 100 µm in size. Binding isotherms data were used for the construction of binding isotherms from which binding between pepsin and polystyrene microplastic was described. Structural features of pepsin in the presence of polystyrene were analysed by CD spectrometry and spectrofluorimetry.",
title = "Research data no. 1 for the manuscript: Ovalbumin interaction with polystyrene and polyethylene terephthalate microplastics alters its structural properties",
url = "https://hdl.handle.net/21.15107/rcub_cherry_6465"
}

Gligorijević, N., Lujić, T., Mutić, T., Vasović, T., de Guzman, M. K., Aćimović, J., Stanić-Vučinić, D.,& Ćirković-Veličković, T.. (2024). Research data no. 1 for the manuscript: Ovalbumin interaction with polystyrene and polyethylene terephthalate microplastics alters its structural properties. .
https://hdl.handle.net/21.15107/rcub_cherry_6465

Gligorijević N, Lujić T, Mutić T, Vasović T, de Guzman MK, Aćimović J, Stanić-Vučinić D, Ćirković-Veličković T. Research data no. 1 for the manuscript: Ovalbumin interaction with polystyrene and polyethylene terephthalate microplastics alters its structural properties. 2024;.
https://hdl.handle.net/21.15107/rcub_cherry_6465 .

Gligorijević, Nikola, Lujić, Tamara, Mutić, Tamara, Vasović, Tamara, de Guzman, Maria Krishna, Aćimović, Jelena, Stanić-Vučinić, Dragana, Ćirković-Veličković, Tanja, "Research data no. 1 for the manuscript: Ovalbumin interaction with polystyrene and polyethylene terephthalate microplastics alters its structural properties" (2024),
https://hdl.handle.net/21.15107/rcub_cherry_6465 .

TY  - JOUR
AU  - Khulal, Urmila
AU  - Đukić, Teodora
AU  - Smiljanić, Katarina
AU  - Vasović, Tamara
AU  - Aćimović, Jelena
AU  - Rajković, Andreja
AU  - Ćirković-Veličković, Tanja
PY  - 2024
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6437
AB  - Meat samples were subjected to thermal processing combined with simulated INFOGEST in vitro gastrointestinal (GI) digestion. Protein modifications (PMs) were screened with commercially available PM-specific antibodies. Specific proteins at 20, 37, 50, and 65 kDa react to more than 3 PM-specific antibodies among meat proteins. Lysine methylation and methionine oxidation were the most prominent PMs in WB. Mass spectrometry confirmed bands at ≈20 kDa as allergenic proteins: sarcoplasmic calcium-binding protein in oyster, 37 kDa as tropomyosin in shrimp, oyster, and abalone. MS-identified PMs of shellfish allergens were aligned to the IgE binding epitopes. GI digestion-resistant peptides of shellfish proteins were identified as paramyosins in oyster and abalone and SBP in shrimp. Our results point to the high susceptibility of immunodominant epitopes of major shellfish allergens to PMs. In TPM, saturation of oxidative modification increases with thermal processing resulting in higher susceptibility of TPM to gastric digestion.
PB  - Elsevier
T2  - Journal of Functional Foods
T1  - Protein modifications screening of raw and thermally treated meat gastrointestinal digesta
VL  - 113
SP  - 106052
DO  - 10.1016/j.jff.2024.106052
ER  - 

@article{
author = "Khulal, Urmila and Đukić, Teodora and Smiljanić, Katarina and Vasović, Tamara and Aćimović, Jelena and Rajković, Andreja and Ćirković-Veličković, Tanja",
year = "2024",
abstract = "Meat samples were subjected to thermal processing combined with simulated INFOGEST in vitro gastrointestinal (GI) digestion. Protein modifications (PMs) were screened with commercially available PM-specific antibodies. Specific proteins at 20, 37, 50, and 65 kDa react to more than 3 PM-specific antibodies among meat proteins. Lysine methylation and methionine oxidation were the most prominent PMs in WB. Mass spectrometry confirmed bands at ≈20 kDa as allergenic proteins: sarcoplasmic calcium-binding protein in oyster, 37 kDa as tropomyosin in shrimp, oyster, and abalone. MS-identified PMs of shellfish allergens were aligned to the IgE binding epitopes. GI digestion-resistant peptides of shellfish proteins were identified as paramyosins in oyster and abalone and SBP in shrimp. Our results point to the high susceptibility of immunodominant epitopes of major shellfish allergens to PMs. In TPM, saturation of oxidative modification increases with thermal processing resulting in higher susceptibility of TPM to gastric digestion.",
publisher = "Elsevier",
journal = "Journal of Functional Foods",
title = "Protein modifications screening of raw and thermally treated meat gastrointestinal digesta",
volume = "113",
pages = "106052",
doi = "10.1016/j.jff.2024.106052"
}

Khulal, U., Đukić, T., Smiljanić, K., Vasović, T., Aćimović, J., Rajković, A.,& Ćirković-Veličković, T.. (2024). Protein modifications screening of raw and thermally treated meat gastrointestinal digesta. in Journal of Functional Foods
Elsevier., 113, 106052.
https://doi.org/10.1016/j.jff.2024.106052

Khulal U, Đukić T, Smiljanić K, Vasović T, Aćimović J, Rajković A, Ćirković-Veličković T. Protein modifications screening of raw and thermally treated meat gastrointestinal digesta. in Journal of Functional Foods. 2024;113:106052.
doi:10.1016/j.jff.2024.106052 .

Khulal, Urmila, Đukić, Teodora, Smiljanić, Katarina, Vasović, Tamara, Aćimović, Jelena, Rajković, Andreja, Ćirković-Veličković, Tanja, "Protein modifications screening of raw and thermally treated meat gastrointestinal digesta" in Journal of Functional Foods, 113 (2024):106052,
https://doi.org/10.1016/j.jff.2024.106052 . .

TY  - JOUR
AU  - Krishna de Guzman, Maria
AU  - Stanić-Vučinić, Dragana
AU  - Gligorijević, Nikola
AU  - Wimmer, Lukas
AU  - Gasparyan, Manvel
AU  - Lujić, Tamara
AU  - Vasović, Tamara
AU  - Dailey, Lea Ann
AU  - Van Haute, Sam
AU  - Ćirković-Veličković, Tanja
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6320
AB  - Human ingestion of microplastics (MPs) is common and inevitable due to the widespread contamination of food items, but implications on the gastric digestion of food proteins are still unknown. In this study, the interactions between pepsin and polystyrene (PS) MPs were evaluated by investigating enzyme activity and conformation in a simulated human gastric environment in the presence or absence of PS MPs. The impact on food digestion was also assessed by monitoring the kinetics of protein hydrolysis through static in vitro gastric digestion of cow's milk contaminated with PS. The binding of pepsin to PS showed that the surface chemistry of MPs dictates binding affinity. The key contributor to pepsin adsorption seems to be π−π interactions between the aromatic residues and the PS phenyl rings. During quick exposure (10 min) of pepsin to increasing concentrations (222, 2219, 22188 particles/mL) of 10 μm PS (PS10) and 100 μm PS (PS100), total enzymatic activities were not affected remarkably. However, upon prolonged exposure at 1 and 2 h, preferential binding of pepsin to the small, low zeta-potential PS caused structural changes in the protein which led to a significant reduction of its activity. Digestion of cow's milk mixed with PS10 resulted in transient accumulation of larger peptides (10–35 kDa) and reduced bioavailability of short peptides (2–9 kDa) in the gastric phase. This, however, was only observed at extremely high PS10 concentration (0.3 mg/mL or 5.46E+05 particles/mL). The digestion of milk peptides, bound preferentially over pepsin within the hard corona on the PS10 surface, was delayed up to 15 min in comparison to bulk protein digestion. Intact caseins, otherwise rapidly digested, remained bound to PS10 in the hard corona for up to 15 min. This work presents valuable insights regarding the interaction of MPs, food proteins, and pepsin, and their dynamics during gastric digestion.
PB  - Elsevier
T2  - Environmental Pollution
T1  - Small polystyrene microplastics interfere with the breakdown of milk proteins during static in vitro simulated human gastric digestion
VL  - 335
SP  - 122282
DO  - 10.1016/j.envpol.2023.122282
ER  - 

@article{
author = "Krishna de Guzman, Maria and Stanić-Vučinić, Dragana and Gligorijević, Nikola and Wimmer, Lukas and Gasparyan, Manvel and Lujić, Tamara and Vasović, Tamara and Dailey, Lea Ann and Van Haute, Sam and Ćirković-Veličković, Tanja",
year = "2023",
abstract = "Human ingestion of microplastics (MPs) is common and inevitable due to the widespread contamination of food items, but implications on the gastric digestion of food proteins are still unknown. In this study, the interactions between pepsin and polystyrene (PS) MPs were evaluated by investigating enzyme activity and conformation in a simulated human gastric environment in the presence or absence of PS MPs. The impact on food digestion was also assessed by monitoring the kinetics of protein hydrolysis through static in vitro gastric digestion of cow's milk contaminated with PS. The binding of pepsin to PS showed that the surface chemistry of MPs dictates binding affinity. The key contributor to pepsin adsorption seems to be π−π interactions between the aromatic residues and the PS phenyl rings. During quick exposure (10 min) of pepsin to increasing concentrations (222, 2219, 22188 particles/mL) of 10 μm PS (PS10) and 100 μm PS (PS100), total enzymatic activities were not affected remarkably. However, upon prolonged exposure at 1 and 2 h, preferential binding of pepsin to the small, low zeta-potential PS caused structural changes in the protein which led to a significant reduction of its activity. Digestion of cow's milk mixed with PS10 resulted in transient accumulation of larger peptides (10–35 kDa) and reduced bioavailability of short peptides (2–9 kDa) in the gastric phase. This, however, was only observed at extremely high PS10 concentration (0.3 mg/mL or 5.46E+05 particles/mL). The digestion of milk peptides, bound preferentially over pepsin within the hard corona on the PS10 surface, was delayed up to 15 min in comparison to bulk protein digestion. Intact caseins, otherwise rapidly digested, remained bound to PS10 in the hard corona for up to 15 min. This work presents valuable insights regarding the interaction of MPs, food proteins, and pepsin, and their dynamics during gastric digestion.",
publisher = "Elsevier",
journal = "Environmental Pollution",
title = "Small polystyrene microplastics interfere with the breakdown of milk proteins during static in vitro simulated human gastric digestion",
volume = "335",
pages = "122282",
doi = "10.1016/j.envpol.2023.122282"
}

Krishna de Guzman, M., Stanić-Vučinić, D., Gligorijević, N., Wimmer, L., Gasparyan, M., Lujić, T., Vasović, T., Dailey, L. A., Van Haute, S.,& Ćirković-Veličković, T.. (2023). Small polystyrene microplastics interfere with the breakdown of milk proteins during static in vitro simulated human gastric digestion. in Environmental Pollution
Elsevier., 335, 122282.
https://doi.org/10.1016/j.envpol.2023.122282

Krishna de Guzman M, Stanić-Vučinić D, Gligorijević N, Wimmer L, Gasparyan M, Lujić T, Vasović T, Dailey LA, Van Haute S, Ćirković-Veličković T. Small polystyrene microplastics interfere with the breakdown of milk proteins during static in vitro simulated human gastric digestion. in Environmental Pollution. 2023;335:122282.
doi:10.1016/j.envpol.2023.122282 .

Krishna de Guzman, Maria, Stanić-Vučinić, Dragana, Gligorijević, Nikola, Wimmer, Lukas, Gasparyan, Manvel, Lujić, Tamara, Vasović, Tamara, Dailey, Lea Ann, Van Haute, Sam, Ćirković-Veličković, Tanja, "Small polystyrene microplastics interfere with the breakdown of milk proteins during static in vitro simulated human gastric digestion" in Environmental Pollution, 335 (2023):122282,
https://doi.org/10.1016/j.envpol.2023.122282 . .

TY  - DATA
AU  - de Guzman, Maria Krishna
AU  - Stanić-Vučinić, Dragana
AU  - Gligorijević, Nikola
AU  - Wimmer, Lukas
AU  - Gasparyan, Manvel
AU  - Lujić, Tamara
AU  - Vasović, Tamara
AU  - Dailey, Lea Ann
AU  - Van Haute, Sam
AU  - Ćirković-Veličković, Tanja
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6463
AB  - The analysis of structural changes of pepsin in the presence of polystyrene microplastic of 10 and 100 µm in size in simulated gastric fluid. Data obtained from J-815 CD spectropolarimeter, and further analyzed by Origin and Excel software.
T2  - Environmental Pollution
T1  - Research data for: Krishna de Guzman, M., Stanić-Vučinić, D., Gligorijević, N., Wimmer, L., Gasparyan, M., Lujić, T., Vasović, T., Dailey, L. A., Van Haute, S.,& Ćirković-Veličković, T.. (2023). Small polystyrene microplastics interfere with the breakdown of milk proteins during static in vitro simulated human gastric digestion. in Environmental Pollution Elsevier., 335, 122282. https://doi.org/10.1016/j.envpol.2023.122282
UR  - https://hdl.handle.net/21.15107/rcub_cherry_6463
ER  - 

@misc{
author = "de Guzman, Maria Krishna and Stanić-Vučinić, Dragana and Gligorijević, Nikola and Wimmer, Lukas and Gasparyan, Manvel and Lujić, Tamara and Vasović, Tamara and Dailey, Lea Ann and Van Haute, Sam and Ćirković-Veličković, Tanja",
year = "2023",
abstract = "The analysis of structural changes of pepsin in the presence of polystyrene microplastic of 10 and 100 µm in size in simulated gastric fluid. Data obtained from J-815 CD spectropolarimeter, and further analyzed by Origin and Excel software.",
journal = "Environmental Pollution",
title = "Research data for: Krishna de Guzman, M., Stanić-Vučinić, D., Gligorijević, N., Wimmer, L., Gasparyan, M., Lujić, T., Vasović, T., Dailey, L. A., Van Haute, S.,& Ćirković-Veličković, T.. (2023). Small polystyrene microplastics interfere with the breakdown of milk proteins during static in vitro simulated human gastric digestion. in Environmental Pollution Elsevier., 335, 122282. https://doi.org/10.1016/j.envpol.2023.122282",
url = "https://hdl.handle.net/21.15107/rcub_cherry_6463"
}

de Guzman, M. K., Stanić-Vučinić, D., Gligorijević, N., Wimmer, L., Gasparyan, M., Lujić, T., Vasović, T., Dailey, L. A., Van Haute, S.,& Ćirković-Veličković, T.. (2023). Research data for: Krishna de Guzman, M., Stanić-Vučinić, D., Gligorijević, N., Wimmer, L., Gasparyan, M., Lujić, T., Vasović, T., Dailey, L. A., Van Haute, S.,& Ćirković-Veličković, T.. (2023). Small polystyrene microplastics interfere with the breakdown of milk proteins during static in vitro simulated human gastric digestion. in Environmental Pollution Elsevier., 335, 122282. https://doi.org/10.1016/j.envpol.2023.122282. in Environmental Pollution.
https://hdl.handle.net/21.15107/rcub_cherry_6463

de Guzman MK, Stanić-Vučinić D, Gligorijević N, Wimmer L, Gasparyan M, Lujić T, Vasović T, Dailey LA, Van Haute S, Ćirković-Veličković T. Research data for: Krishna de Guzman, M., Stanić-Vučinić, D., Gligorijević, N., Wimmer, L., Gasparyan, M., Lujić, T., Vasović, T., Dailey, L. A., Van Haute, S.,& Ćirković-Veličković, T.. (2023). Small polystyrene microplastics interfere with the breakdown of milk proteins during static in vitro simulated human gastric digestion. in Environmental Pollution Elsevier., 335, 122282. https://doi.org/10.1016/j.envpol.2023.122282. in Environmental Pollution. 2023;.
https://hdl.handle.net/21.15107/rcub_cherry_6463 .

de Guzman, Maria Krishna, Stanić-Vučinić, Dragana, Gligorijević, Nikola, Wimmer, Lukas, Gasparyan, Manvel, Lujić, Tamara, Vasović, Tamara, Dailey, Lea Ann, Van Haute, Sam, Ćirković-Veličković, Tanja, "Research data for: Krishna de Guzman, M., Stanić-Vučinić, D., Gligorijević, N., Wimmer, L., Gasparyan, M., Lujić, T., Vasović, T., Dailey, L. A., Van Haute, S.,& Ćirković-Veličković, T.. (2023). Small polystyrene microplastics interfere with the breakdown of milk proteins during static in vitro simulated human gastric digestion. in Environmental Pollution Elsevier., 335, 122282. https://doi.org/10.1016/j.envpol.2023.122282" in Environmental Pollution (2023),
https://hdl.handle.net/21.15107/rcub_cherry_6463 .

TY  - DATA
AU  - Đapović, Milica
AU  - Apostolović, Danijela
AU  - Poštić, Vojislava
AU  - Lujić, Tamara
AU  - Jovanović, Vesna
AU  - Stanić-Vučinić, Dragana
AU  - van Hage, Marianne
AU  - Maslak, Veselin
AU  - Ćirković-Veličković, Tanja
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6464
AB  - Determination of hydrodynamic diameter of PET NPs washed and dispersed in water, BSA (0.05%), and SDS (0.1%), and NPs unwashed and dispersed in water.  Zeta potential, mobility, and conductivity determination in PET NPs at different stages of purification. Output from the Malvern zetasizer Nano-ZS ZEN 3600 (Malvern Panalytical, Malvern, UK).
PB  - MDPI
T2  - Polymers
T1  - Research data no. 2 for: Djapovic, M., Apostolović, D., Postic, V., Lujić, T., Jovanović, V., Stanić-Vučinić, D., van Hage, M., Maslak, V.,& Ćirković-Veličković, T.. (2023). Characterization of Nanoprecipitated PET Nanoplastics by 1H NMR and Impact of Residual Ionic Surfactant on Viability of Human Primary Mononuclear Cells and Hemolysis of Erythrocytes. in Polymers MDPI., 15(24), 4703. https://doi.org/10.3390/polym15244703
UR  - https://hdl.handle.net/21.15107/rcub_cherry_6464
ER  - 

@misc{
author = "Đapović, Milica and Apostolović, Danijela and Poštić, Vojislava and Lujić, Tamara and Jovanović, Vesna and Stanić-Vučinić, Dragana and van Hage, Marianne and Maslak, Veselin and Ćirković-Veličković, Tanja",
year = "2023",
abstract = "Determination of hydrodynamic diameter of PET NPs washed and dispersed in water, BSA (0.05%), and SDS (0.1%), and NPs unwashed and dispersed in water.  Zeta potential, mobility, and conductivity determination in PET NPs at different stages of purification. Output from the Malvern zetasizer Nano-ZS ZEN 3600 (Malvern Panalytical, Malvern, UK).",
publisher = "MDPI",
journal = "Polymers",
title = "Research data no. 2 for: Djapovic, M., Apostolović, D., Postic, V., Lujić, T., Jovanović, V., Stanić-Vučinić, D., van Hage, M., Maslak, V.,& Ćirković-Veličković, T.. (2023). Characterization of Nanoprecipitated PET Nanoplastics by 1H NMR and Impact of Residual Ionic Surfactant on Viability of Human Primary Mononuclear Cells and Hemolysis of Erythrocytes. in Polymers MDPI., 15(24), 4703. https://doi.org/10.3390/polym15244703",
url = "https://hdl.handle.net/21.15107/rcub_cherry_6464"
}

Đapović, M., Apostolović, D., Poštić, V., Lujić, T., Jovanović, V., Stanić-Vučinić, D., van Hage, M., Maslak, V.,& Ćirković-Veličković, T.. (2023). Research data no. 2 for: Djapovic, M., Apostolović, D., Postic, V., Lujić, T., Jovanović, V., Stanić-Vučinić, D., van Hage, M., Maslak, V.,& Ćirković-Veličković, T.. (2023). Characterization of Nanoprecipitated PET Nanoplastics by 1H NMR and Impact of Residual Ionic Surfactant on Viability of Human Primary Mononuclear Cells and Hemolysis of Erythrocytes. in Polymers MDPI., 15(24), 4703. https://doi.org/10.3390/polym15244703. in Polymers
MDPI..
https://hdl.handle.net/21.15107/rcub_cherry_6464

Đapović M, Apostolović D, Poštić V, Lujić T, Jovanović V, Stanić-Vučinić D, van Hage M, Maslak V, Ćirković-Veličković T. Research data no. 2 for: Djapovic, M., Apostolović, D., Postic, V., Lujić, T., Jovanović, V., Stanić-Vučinić, D., van Hage, M., Maslak, V.,& Ćirković-Veličković, T.. (2023). Characterization of Nanoprecipitated PET Nanoplastics by 1H NMR and Impact of Residual Ionic Surfactant on Viability of Human Primary Mononuclear Cells and Hemolysis of Erythrocytes. in Polymers MDPI., 15(24), 4703. https://doi.org/10.3390/polym15244703. in Polymers. 2023;.
https://hdl.handle.net/21.15107/rcub_cherry_6464 .

Đapović, Milica, Apostolović, Danijela, Poštić, Vojislava, Lujić, Tamara, Jovanović, Vesna, Stanić-Vučinić, Dragana, van Hage, Marianne, Maslak, Veselin, Ćirković-Veličković, Tanja, "Research data no. 2 for: Djapovic, M., Apostolović, D., Postic, V., Lujić, T., Jovanović, V., Stanić-Vučinić, D., van Hage, M., Maslak, V.,& Ćirković-Veličković, T.. (2023). Characterization of Nanoprecipitated PET Nanoplastics by 1H NMR and Impact of Residual Ionic Surfactant on Viability of Human Primary Mononuclear Cells and Hemolysis of Erythrocytes. in Polymers MDPI., 15(24), 4703. https://doi.org/10.3390/polym15244703" in Polymers (2023),
https://hdl.handle.net/21.15107/rcub_cherry_6464 .

TY  - DATA
AU  - Đapović, Milica
AU  - Apostolović, Danijela
AU  - Poštić, Vojislava
AU  - Lujić, Tamara
AU  - Jovanović, Vesna
AU  - Stanić-Vučinić, Dragana
AU  - van Hage, Marianne
AU  - Maslak, Veselin
AU  - Ćirković-Veličković, Tanja
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6467
AB  - The presence of the ionic surfactant sodium dodecyl sulfate (SDS) was characterized by 1H NMR, where the relative ratio of NP/surfactant was monitored on the basis of the chemical shifts characteristic of PET and SDS. 1H NMR spectra of the NPs preparation before and during all the washing steps. Determination of SDS Level in Corona of PET NPs by 1H NMR. Output from the Varian/Agilent NMR 400 MHz. NMR spectra were processed in Mnova software.
PB  - MDPI
T2  - Polymers
T1  - Research data no. 3 for: Djapovic, M., Apostolović, D., Postic, V., Lujić, T., Jovanović, V., Stanić-Vučinić, D., van Hage, M., Maslak, V.,& Ćirković-Veličković, T.. (2023). Characterization of Nanoprecipitated PET Nanoplastics by 1H NMR and Impact of Residual Ionic Surfactant on Viability of Human Primary Mononuclear Cells and Hemolysis of Erythrocytes. in Polymers MDPI., 15(24), 4703. https://doi.org/10.3390/polym15244703
UR  - https://hdl.handle.net/21.15107/rcub_cherry_6467
ER  - 

@misc{
author = "Đapović, Milica and Apostolović, Danijela and Poštić, Vojislava and Lujić, Tamara and Jovanović, Vesna and Stanić-Vučinić, Dragana and van Hage, Marianne and Maslak, Veselin and Ćirković-Veličković, Tanja",
year = "2023",
abstract = "The presence of the ionic surfactant sodium dodecyl sulfate (SDS) was characterized by 1H NMR, where the relative ratio of NP/surfactant was monitored on the basis of the chemical shifts characteristic of PET and SDS. 1H NMR spectra of the NPs preparation before and during all the washing steps. Determination of SDS Level in Corona of PET NPs by 1H NMR. Output from the Varian/Agilent NMR 400 MHz. NMR spectra were processed in Mnova software.",
publisher = "MDPI",
journal = "Polymers",
title = "Research data no. 3 for: Djapovic, M., Apostolović, D., Postic, V., Lujić, T., Jovanović, V., Stanić-Vučinić, D., van Hage, M., Maslak, V.,& Ćirković-Veličković, T.. (2023). Characterization of Nanoprecipitated PET Nanoplastics by 1H NMR and Impact of Residual Ionic Surfactant on Viability of Human Primary Mononuclear Cells and Hemolysis of Erythrocytes. in Polymers MDPI., 15(24), 4703. https://doi.org/10.3390/polym15244703",
url = "https://hdl.handle.net/21.15107/rcub_cherry_6467"
}

Đapović, M., Apostolović, D., Poštić, V., Lujić, T., Jovanović, V., Stanić-Vučinić, D., van Hage, M., Maslak, V.,& Ćirković-Veličković, T.. (2023). Research data no. 3 for: Djapovic, M., Apostolović, D., Postic, V., Lujić, T., Jovanović, V., Stanić-Vučinić, D., van Hage, M., Maslak, V.,& Ćirković-Veličković, T.. (2023). Characterization of Nanoprecipitated PET Nanoplastics by 1H NMR and Impact of Residual Ionic Surfactant on Viability of Human Primary Mononuclear Cells and Hemolysis of Erythrocytes. in Polymers MDPI., 15(24), 4703. https://doi.org/10.3390/polym15244703. in Polymers
MDPI..
https://hdl.handle.net/21.15107/rcub_cherry_6467

Đapović M, Apostolović D, Poštić V, Lujić T, Jovanović V, Stanić-Vučinić D, van Hage M, Maslak V, Ćirković-Veličković T. Research data no. 3 for: Djapovic, M., Apostolović, D., Postic, V., Lujić, T., Jovanović, V., Stanić-Vučinić, D., van Hage, M., Maslak, V.,& Ćirković-Veličković, T.. (2023). Characterization of Nanoprecipitated PET Nanoplastics by 1H NMR and Impact of Residual Ionic Surfactant on Viability of Human Primary Mononuclear Cells and Hemolysis of Erythrocytes. in Polymers MDPI., 15(24), 4703. https://doi.org/10.3390/polym15244703. in Polymers. 2023;.
https://hdl.handle.net/21.15107/rcub_cherry_6467 .

Đapović, Milica, Apostolović, Danijela, Poštić, Vojislava, Lujić, Tamara, Jovanović, Vesna, Stanić-Vučinić, Dragana, van Hage, Marianne, Maslak, Veselin, Ćirković-Veličković, Tanja, "Research data no. 3 for: Djapovic, M., Apostolović, D., Postic, V., Lujić, T., Jovanović, V., Stanić-Vučinić, D., van Hage, M., Maslak, V.,& Ćirković-Veličković, T.. (2023). Characterization of Nanoprecipitated PET Nanoplastics by 1H NMR and Impact of Residual Ionic Surfactant on Viability of Human Primary Mononuclear Cells and Hemolysis of Erythrocytes. in Polymers MDPI., 15(24), 4703. https://doi.org/10.3390/polym15244703" in Polymers (2023),
https://hdl.handle.net/21.15107/rcub_cherry_6467 .

TY  - DATA
AU  - Đapović, Milica
AU  - Apostolović, Danijela
AU  - Poštić, Vojislava
AU  - Lujić, Tamara
AU  - Jovanović, Vesna
AU  - Stanić-Vučinić, Dragana
AU  - van Hage, Marianne
AU  - Maslak, Veselin
AU  - Ćirković-Veličković, Tanja
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6468
AB  - Size distributions (by intensity percentage) of NPs washed suspended in different dispersants, water,BSA (0.05%),SDS (0.1 %). Distributions data from different combined fractions: from NP-10 to NP-30; from NP-40 to NP-60 and from NP-70 to NP-90. Output from the Malvern zetasizer Nano-ZS ZEN 3600 (Malvern Panalytical, Malvern, UK).
PB  - MDPI
T2  - Polymers
T1  - Research data no. 4 for: Djapovic, M., Apostolović, D., Postic, V., Lujić, T., Jovanović, V., Stanić-Vučinić, D., van Hage, M., Maslak, V.,& Ćirković-Veličković, T.. (2023). Characterization of Nanoprecipitated PET Nanoplastics by 1H NMR and Impact of Residual Ionic Surfactant on Viability of Human Primary Mononuclear Cells and Hemolysis of Erythrocytes. in Polymers MDPI., 15(24), 4703. https://doi.org/10.3390/polym15244703
UR  - https://hdl.handle.net/21.15107/rcub_cherry_6468
ER  - 

@misc{
author = "Đapović, Milica and Apostolović, Danijela and Poštić, Vojislava and Lujić, Tamara and Jovanović, Vesna and Stanić-Vučinić, Dragana and van Hage, Marianne and Maslak, Veselin and Ćirković-Veličković, Tanja",
year = "2023",
abstract = "Size distributions (by intensity percentage) of NPs washed suspended in different dispersants, water,BSA (0.05%),SDS (0.1 %). Distributions data from different combined fractions: from NP-10 to NP-30; from NP-40 to NP-60 and from NP-70 to NP-90. Output from the Malvern zetasizer Nano-ZS ZEN 3600 (Malvern Panalytical, Malvern, UK).",
publisher = "MDPI",
journal = "Polymers",
title = "Research data no. 4 for: Djapovic, M., Apostolović, D., Postic, V., Lujić, T., Jovanović, V., Stanić-Vučinić, D., van Hage, M., Maslak, V.,& Ćirković-Veličković, T.. (2023). Characterization of Nanoprecipitated PET Nanoplastics by 1H NMR and Impact of Residual Ionic Surfactant on Viability of Human Primary Mononuclear Cells and Hemolysis of Erythrocytes. in Polymers MDPI., 15(24), 4703. https://doi.org/10.3390/polym15244703",
url = "https://hdl.handle.net/21.15107/rcub_cherry_6468"
}

Đapović, M., Apostolović, D., Poštić, V., Lujić, T., Jovanović, V., Stanić-Vučinić, D., van Hage, M., Maslak, V.,& Ćirković-Veličković, T.. (2023). Research data no. 4 for: Djapovic, M., Apostolović, D., Postic, V., Lujić, T., Jovanović, V., Stanić-Vučinić, D., van Hage, M., Maslak, V.,& Ćirković-Veličković, T.. (2023). Characterization of Nanoprecipitated PET Nanoplastics by 1H NMR and Impact of Residual Ionic Surfactant on Viability of Human Primary Mononuclear Cells and Hemolysis of Erythrocytes. in Polymers MDPI., 15(24), 4703. https://doi.org/10.3390/polym15244703. in Polymers
MDPI..
https://hdl.handle.net/21.15107/rcub_cherry_6468

Đapović M, Apostolović D, Poštić V, Lujić T, Jovanović V, Stanić-Vučinić D, van Hage M, Maslak V, Ćirković-Veličković T. Research data no. 4 for: Djapovic, M., Apostolović, D., Postic, V., Lujić, T., Jovanović, V., Stanić-Vučinić, D., van Hage, M., Maslak, V.,& Ćirković-Veličković, T.. (2023). Characterization of Nanoprecipitated PET Nanoplastics by 1H NMR and Impact of Residual Ionic Surfactant on Viability of Human Primary Mononuclear Cells and Hemolysis of Erythrocytes. in Polymers MDPI., 15(24), 4703. https://doi.org/10.3390/polym15244703. in Polymers. 2023;.
https://hdl.handle.net/21.15107/rcub_cherry_6468 .

Đapović, Milica, Apostolović, Danijela, Poštić, Vojislava, Lujić, Tamara, Jovanović, Vesna, Stanić-Vučinić, Dragana, van Hage, Marianne, Maslak, Veselin, Ćirković-Veličković, Tanja, "Research data no. 4 for: Djapovic, M., Apostolović, D., Postic, V., Lujić, T., Jovanović, V., Stanić-Vučinić, D., van Hage, M., Maslak, V.,& Ćirković-Veličković, T.. (2023). Characterization of Nanoprecipitated PET Nanoplastics by 1H NMR and Impact of Residual Ionic Surfactant on Viability of Human Primary Mononuclear Cells and Hemolysis of Erythrocytes. in Polymers MDPI., 15(24), 4703. https://doi.org/10.3390/polym15244703" in Polymers (2023),
https://hdl.handle.net/21.15107/rcub_cherry_6468 .

TY  - JOUR
AU  - Djapovic, Milica
AU  - Apostolović, Danijela
AU  - Postic, Vojislava
AU  - Lujić, Tamara
AU  - Jovanović, Vesna
AU  - Stanić-Vučinić, Dragana
AU  - van Hage, Marianne
AU  - Maslak, Veselin
AU  - Ćirković-Veličković, Tanja
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6433
AB  - Manufactured nanoplastic particles (NPs) are indispensable for in vitro and in vivo testing and a health risk assessment of this emerging environmental contaminant is needed. The high surface area and inherent hydrophobicity of plastic materials makes the production of NPs devoid of any contaminants very challenging. In this study, we produced nanoprecipitated polyethylene terephthalate (PET) NPs (300 nm hydrodynamic diameter) with an overall yield of 0.76%. The presence of the ionic surfactant sodium dodecyl sulfate (SDS) was characterized by 1H NMR, where the relative ratio of NP/surfactant was monitored on the basis of the chemical shifts characteristic of PET and SDS. For a wide range of surfactant/NP ratios (17:100 to 1.2:100), the measured zeta potential changed from −42.10 to −34.93 mV, but with an NP concentration up to 100 μg/mL, no clear differences were observed in the cellular assays performed in protein-rich media on primary human cells. The remaining impurities contributed to the outcome of the biological assays applied in protein-free buffers, such as human red blood cell hemolysis. The presence of SDS increased the NP-induced hemolysis by 1.5% in protein-rich buffer and by 7.5% in protein-free buffer. As the size, shape, zeta potential, and contaminants of NPs may all be relevant parameters for the biological effects of NPs, the relative quantification of impurities exemplified in our work by the application of 1H NMR for PET NPs and the ionic surfactant SDS could be a valuable auxiliary method in the quality control of manufactured NPs.
PB  - MDPI
T2  - Polymers
T1  - Characterization of Nanoprecipitated PET Nanoplastics by 1H NMR and Impact of Residual Ionic Surfactant on Viability of Human Primary Mononuclear Cells and Hemolysis of Erythrocytes
VL  - 15
IS  - 24
SP  - 4703
DO  - 10.3390/polym15244703
ER  - 

@article{
author = "Djapovic, Milica and Apostolović, Danijela and Postic, Vojislava and Lujić, Tamara and Jovanović, Vesna and Stanić-Vučinić, Dragana and van Hage, Marianne and Maslak, Veselin and Ćirković-Veličković, Tanja",
year = "2023",
abstract = "Manufactured nanoplastic particles (NPs) are indispensable for in vitro and in vivo testing and a health risk assessment of this emerging environmental contaminant is needed. The high surface area and inherent hydrophobicity of plastic materials makes the production of NPs devoid of any contaminants very challenging. In this study, we produced nanoprecipitated polyethylene terephthalate (PET) NPs (300 nm hydrodynamic diameter) with an overall yield of 0.76%. The presence of the ionic surfactant sodium dodecyl sulfate (SDS) was characterized by 1H NMR, where the relative ratio of NP/surfactant was monitored on the basis of the chemical shifts characteristic of PET and SDS. For a wide range of surfactant/NP ratios (17:100 to 1.2:100), the measured zeta potential changed from −42.10 to −34.93 mV, but with an NP concentration up to 100 μg/mL, no clear differences were observed in the cellular assays performed in protein-rich media on primary human cells. The remaining impurities contributed to the outcome of the biological assays applied in protein-free buffers, such as human red blood cell hemolysis. The presence of SDS increased the NP-induced hemolysis by 1.5% in protein-rich buffer and by 7.5% in protein-free buffer. As the size, shape, zeta potential, and contaminants of NPs may all be relevant parameters for the biological effects of NPs, the relative quantification of impurities exemplified in our work by the application of 1H NMR for PET NPs and the ionic surfactant SDS could be a valuable auxiliary method in the quality control of manufactured NPs.",
publisher = "MDPI",
journal = "Polymers",
title = "Characterization of Nanoprecipitated PET Nanoplastics by 1H NMR and Impact of Residual Ionic Surfactant on Viability of Human Primary Mononuclear Cells and Hemolysis of Erythrocytes",
volume = "15",
number = "24",
pages = "4703",
doi = "10.3390/polym15244703"
}

Djapovic, M., Apostolović, D., Postic, V., Lujić, T., Jovanović, V., Stanić-Vučinić, D., van Hage, M., Maslak, V.,& Ćirković-Veličković, T.. (2023). Characterization of Nanoprecipitated PET Nanoplastics by 1H NMR and Impact of Residual Ionic Surfactant on Viability of Human Primary Mononuclear Cells and Hemolysis of Erythrocytes. in Polymers
MDPI., 15(24), 4703.
https://doi.org/10.3390/polym15244703

Djapovic M, Apostolović D, Postic V, Lujić T, Jovanović V, Stanić-Vučinić D, van Hage M, Maslak V, Ćirković-Veličković T. Characterization of Nanoprecipitated PET Nanoplastics by 1H NMR and Impact of Residual Ionic Surfactant on Viability of Human Primary Mononuclear Cells and Hemolysis of Erythrocytes. in Polymers. 2023;15(24):4703.
doi:10.3390/polym15244703 .

Djapovic, Milica, Apostolović, Danijela, Postic, Vojislava, Lujić, Tamara, Jovanović, Vesna, Stanić-Vučinić, Dragana, van Hage, Marianne, Maslak, Veselin, Ćirković-Veličković, Tanja, "Characterization of Nanoprecipitated PET Nanoplastics by 1H NMR and Impact of Residual Ionic Surfactant on Viability of Human Primary Mononuclear Cells and Hemolysis of Erythrocytes" in Polymers, 15, no. 24 (2023):4703,
https://doi.org/10.3390/polym15244703 . .

TY  - CONF
AU  - Jovanović, Vesna
AU  - Radomirović, Mirjana Ž.
AU  - Krstić-Ristivojević, Maja
AU  - Stanić-Vučinić, Dragana
AU  - Ćirković-Veličković, Tanja
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6023
AB  - In the last several decades, the trend of seafood consumption has significantly increased not only in the countries with a tradition of seafood consumption, but also in other ones [1]. The increase in the world's population and the awareness of healthy food, the globalization of markets, and the development of aquaculture are some of the factors that have led to this trend. The aquaculture of shellfish like clams, mussels, oysters and scallops has been very developed all around the world and the food products based on them have become part of the daily diet for many consumers. In addition, these food products are considered healthy food because of the high content of proteins and essential fatty acids, but their consumption may carry some risks of food allergy. Tropomyosin from shellfish (TPM) is the major allergen responsible for the development of anaphylaxis in persons with food allergy. The content of TPM in shellfish and its bioavailability from food products can have potential influence on the sensitization of consumers to TPM. It is known that food processing can change the bioavailability of food allergens [2]. The main goal of this study was the investigation of how processing and packaging of shellfish samples can affect the content of TPM in them.
For this study, clam Venerupis philippinarum was chosen as the species with the highest world aquaculture production [1]. After the purchasing of live clams, the animals were separated into 5 groups for the next treatments: fresh live (control group), freshly removed inner content was kept at +4°C for 3 days (three days` shelf-life), frozen in a plastic bag and kept at -20 °C during 7 days, marinated and kept in a glass jar at room temperature during 8 days and freshly boiled. After processing and packaging of samples, the total protein extracts were prepared in 10 mM sodium phosphate buffer pH 7.4 1M NaCl, 1 mM PMSF and the concentration of total proteins was determined by BCA method. The concentration of TPM in the total protein extracts was determined using a sandwich Enzyme-Linked Immunosorbent Assay (ELISA) using in-house prepared clams` TPM standard. The content of TPM (μg) in the samples was expressed per mg of extracted soluble proteins, individual animal and grams of soft wet tissue.
The cooked samples have significantly higher TPM content expressed per gram of soft wet tissue compared to all other treatments. Food processing such as freezing, marinating, or extending the shelf-life at 4°C by 3 days has very little effect on the change in TPM content per gram of soft wet tissue compared to the fresh samples. The processing of clams, like cooking or marinating, caused the content of total soluble extracted proteins to be three to four times lower compared to the other three treatments. In these samples the obtained ratio of the total TPM/ total soluble extracted proteins ratio was the highest. This result can be explained by the fact that TPM is thermostable and stays soluble after cooking, while other proteins become insoluble because of denaturation. The lower ratio of TPM/ total soluble extracted proteins was found in marinated samples compared to the cooked samples. The lowest total tropomyosin/ total soluble extracted proteins ratio was found in 3 days’ shelf-life. Treatments like cooking, marinating and keeping the inner content of the shell at +4°C can significantly affect extractability of proteins, particularly affecting the ratio of major allergen TPM in the total protein extracts. Further studies are needed to examine bioaccessibility of TPM in different biologically relevant fluids (gastric/intestinal) and during digestion in relation to the processing conditions.
PB  - Beograd : Srpsko hemijsko društvo
C3  - XXII EuroFoodChem conference, Book of Abstracts, 14th-16th June, 2023.
T1  - The effect of food processing and packaging of clams on the content of tropomyosin
SP  - 240
EP  - 240
UR  - https://hdl.handle.net/21.15107/rcub_cherry_6023
ER  - 

@conference{
author = "Jovanović, Vesna and Radomirović, Mirjana Ž. and Krstić-Ristivojević, Maja and Stanić-Vučinić, Dragana and Ćirković-Veličković, Tanja",
year = "2023",
abstract = "In the last several decades, the trend of seafood consumption has significantly increased not only in the countries with a tradition of seafood consumption, but also in other ones [1]. The increase in the world's population and the awareness of healthy food, the globalization of markets, and the development of aquaculture are some of the factors that have led to this trend. The aquaculture of shellfish like clams, mussels, oysters and scallops has been very developed all around the world and the food products based on them have become part of the daily diet for many consumers. In addition, these food products are considered healthy food because of the high content of proteins and essential fatty acids, but their consumption may carry some risks of food allergy. Tropomyosin from shellfish (TPM) is the major allergen responsible for the development of anaphylaxis in persons with food allergy. The content of TPM in shellfish and its bioavailability from food products can have potential influence on the sensitization of consumers to TPM. It is known that food processing can change the bioavailability of food allergens [2]. The main goal of this study was the investigation of how processing and packaging of shellfish samples can affect the content of TPM in them.
For this study, clam Venerupis philippinarum was chosen as the species with the highest world aquaculture production [1]. After the purchasing of live clams, the animals were separated into 5 groups for the next treatments: fresh live (control group), freshly removed inner content was kept at +4°C for 3 days (three days` shelf-life), frozen in a plastic bag and kept at -20 °C during 7 days, marinated and kept in a glass jar at room temperature during 8 days and freshly boiled. After processing and packaging of samples, the total protein extracts were prepared in 10 mM sodium phosphate buffer pH 7.4 1M NaCl, 1 mM PMSF and the concentration of total proteins was determined by BCA method. The concentration of TPM in the total protein extracts was determined using a sandwich Enzyme-Linked Immunosorbent Assay (ELISA) using in-house prepared clams` TPM standard. The content of TPM (μg) in the samples was expressed per mg of extracted soluble proteins, individual animal and grams of soft wet tissue.
The cooked samples have significantly higher TPM content expressed per gram of soft wet tissue compared to all other treatments. Food processing such as freezing, marinating, or extending the shelf-life at 4°C by 3 days has very little effect on the change in TPM content per gram of soft wet tissue compared to the fresh samples. The processing of clams, like cooking or marinating, caused the content of total soluble extracted proteins to be three to four times lower compared to the other three treatments. In these samples the obtained ratio of the total TPM/ total soluble extracted proteins ratio was the highest. This result can be explained by the fact that TPM is thermostable and stays soluble after cooking, while other proteins become insoluble because of denaturation. The lower ratio of TPM/ total soluble extracted proteins was found in marinated samples compared to the cooked samples. The lowest total tropomyosin/ total soluble extracted proteins ratio was found in 3 days’ shelf-life. Treatments like cooking, marinating and keeping the inner content of the shell at +4°C can significantly affect extractability of proteins, particularly affecting the ratio of major allergen TPM in the total protein extracts. Further studies are needed to examine bioaccessibility of TPM in different biologically relevant fluids (gastric/intestinal) and during digestion in relation to the processing conditions.",
publisher = "Beograd : Srpsko hemijsko društvo",
journal = "XXII EuroFoodChem conference, Book of Abstracts, 14th-16th June, 2023.",
title = "The effect of food processing and packaging of clams on the content of tropomyosin",
pages = "240-240",
url = "https://hdl.handle.net/21.15107/rcub_cherry_6023"
}

Jovanović, V., Radomirović, M. Ž., Krstić-Ristivojević, M., Stanić-Vučinić, D.,& Ćirković-Veličković, T.. (2023). The effect of food processing and packaging of clams on the content of tropomyosin. in XXII EuroFoodChem conference, Book of Abstracts, 14th-16th June, 2023.
Beograd : Srpsko hemijsko društvo., 240-240.
https://hdl.handle.net/21.15107/rcub_cherry_6023

Jovanović V, Radomirović MŽ, Krstić-Ristivojević M, Stanić-Vučinić D, Ćirković-Veličković T. The effect of food processing and packaging of clams on the content of tropomyosin. in XXII EuroFoodChem conference, Book of Abstracts, 14th-16th June, 2023.. 2023;:240-240.
https://hdl.handle.net/21.15107/rcub_cherry_6023 .

Jovanović, Vesna, Radomirović, Mirjana Ž., Krstić-Ristivojević, Maja, Stanić-Vučinić, Dragana, Ćirković-Veličković, Tanja, "The effect of food processing and packaging of clams on the content of tropomyosin" in XXII EuroFoodChem conference, Book of Abstracts, 14th-16th June, 2023. (2023):240-240,
https://hdl.handle.net/21.15107/rcub_cherry_6023 .

TY  - CONF
AU  - Krstić-Ristivojević, Maja
AU  - Jovanović, Vesna
AU  - Radomirović, Mirjana Ž.
AU  - Trifunović, Olga
AU  - Stanić-Vučinić, Dragana
AU  - Ćirković-Veličković, Tanja
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6024
AB  - In the last 50 years, the annual per capita consumption of seafood products worldwide has more than doubled, from almost 10 kg in 1960 to over 20 kg in 2014. Seafood protein is an essential part of the diet in many countries, particularly where total protein intake is low [1]. However, as defined by the European Community, fish, and shellfish tropomyosins (TPM) are major allergens and major causes of anaphylaxis [2]. The increasing prevalence of food allergies is consistent with the increasing pollution of soil and water with plastic particles. To investigate the potential link between increasing plastic pollution and increasing food allergy prevalence, we aim to develop methods for precise and accurate monitoring of allergens and plastic in real seafood samples.
TPM was isolated from shrimp (Litopenaeus vannamei), clams (Venerupis philippinarum), and mussels (Mytilus galloprovincialis). The obtained in-house TPM proteins from three different sources were resolved using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). The concentration of TPM in seashell samples from different geographical origin was determined using a sandwich Enzyme-Linked Immunosorbent Assay (ELISA) with prior optimization of adequate TPM standard curve using commercial and non-commercial in-house prepared TPM standards.
TPM standards resolved via 2D-PAGE revealed the presence of two isoforms of shrimp and mussels TPM standard, one dominant and one less abundant isoform. Two isoforms from both seafood sources, shrimp and mussels, are slightly different in molecular weight and pI value. As for the TPM standard obtained from clams, the 2D electrophoregram showed possibly eight isoforms with small differences in mass and pI values. Furthermore, the presence of three dominant isoforms can be observed that differ slightly in molecular mass, while other isoforms also differ in pI value. The ELISA results, regarding TPM standard curve optimization, showed that in both the commercial shrimp TPM and in-house shrimp TPM standards, sigmoidal concentration dependence is present in a range of 50 to 0.05 ng/ml, using serial double dilutions. On the other hand, TPM standards isolated from mussels and clams show sigmoidal concentration dependence in the range of 45 to 0.044 μg/ml with using the identical combination of capture and detection antibodies and serial double dilutions. TPM concentrations in clams and mussel samples extrapolated from standard curves of commercial shrimp TPM standard and corresponding in-house TPM standards are presented in Table 1.
Differences in TPM concentration of the same sample using different TPM standards differ from 40 to 600 times, which strongly indicates that the right choice of TPM standard is a critical step for accurate and precise determination of TPM concentration in seafood samples.
PB  - Beograd : Srpsko hemijsko društvo
C3  - XXII EuroFoodChem conference, 14th-16th June, 2023. In: Book of Abstracts
T1  - Tropomyosin quantification in seafood samples-right choice of standard makes a difference
SP  - 132
EP  - 132
UR  - https://hdl.handle.net/21.15107/rcub_cherry_6024
ER  - 

@conference{
author = "Krstić-Ristivojević, Maja and Jovanović, Vesna and Radomirović, Mirjana Ž. and Trifunović, Olga and Stanić-Vučinić, Dragana and Ćirković-Veličković, Tanja",
year = "2023",
abstract = "In the last 50 years, the annual per capita consumption of seafood products worldwide has more than doubled, from almost 10 kg in 1960 to over 20 kg in 2014. Seafood protein is an essential part of the diet in many countries, particularly where total protein intake is low [1]. However, as defined by the European Community, fish, and shellfish tropomyosins (TPM) are major allergens and major causes of anaphylaxis [2]. The increasing prevalence of food allergies is consistent with the increasing pollution of soil and water with plastic particles. To investigate the potential link between increasing plastic pollution and increasing food allergy prevalence, we aim to develop methods for precise and accurate monitoring of allergens and plastic in real seafood samples.
TPM was isolated from shrimp (Litopenaeus vannamei), clams (Venerupis philippinarum), and mussels (Mytilus galloprovincialis). The obtained in-house TPM proteins from three different sources were resolved using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). The concentration of TPM in seashell samples from different geographical origin was determined using a sandwich Enzyme-Linked Immunosorbent Assay (ELISA) with prior optimization of adequate TPM standard curve using commercial and non-commercial in-house prepared TPM standards.
TPM standards resolved via 2D-PAGE revealed the presence of two isoforms of shrimp and mussels TPM standard, one dominant and one less abundant isoform. Two isoforms from both seafood sources, shrimp and mussels, are slightly different in molecular weight and pI value. As for the TPM standard obtained from clams, the 2D electrophoregram showed possibly eight isoforms with small differences in mass and pI values. Furthermore, the presence of three dominant isoforms can be observed that differ slightly in molecular mass, while other isoforms also differ in pI value. The ELISA results, regarding TPM standard curve optimization, showed that in both the commercial shrimp TPM and in-house shrimp TPM standards, sigmoidal concentration dependence is present in a range of 50 to 0.05 ng/ml, using serial double dilutions. On the other hand, TPM standards isolated from mussels and clams show sigmoidal concentration dependence in the range of 45 to 0.044 μg/ml with using the identical combination of capture and detection antibodies and serial double dilutions. TPM concentrations in clams and mussel samples extrapolated from standard curves of commercial shrimp TPM standard and corresponding in-house TPM standards are presented in Table 1.
Differences in TPM concentration of the same sample using different TPM standards differ from 40 to 600 times, which strongly indicates that the right choice of TPM standard is a critical step for accurate and precise determination of TPM concentration in seafood samples.",
publisher = "Beograd : Srpsko hemijsko društvo",
journal = "XXII EuroFoodChem conference, 14th-16th June, 2023. In: Book of Abstracts",
title = "Tropomyosin quantification in seafood samples-right choice of standard makes a difference",
pages = "132-132",
url = "https://hdl.handle.net/21.15107/rcub_cherry_6024"
}

Krstić-Ristivojević, M., Jovanović, V., Radomirović, M. Ž., Trifunović, O., Stanić-Vučinić, D.,& Ćirković-Veličković, T.. (2023). Tropomyosin quantification in seafood samples-right choice of standard makes a difference. in XXII EuroFoodChem conference, 14th-16th June, 2023. In: Book of Abstracts
Beograd : Srpsko hemijsko društvo., 132-132.
https://hdl.handle.net/21.15107/rcub_cherry_6024

Krstić-Ristivojević M, Jovanović V, Radomirović MŽ, Trifunović O, Stanić-Vučinić D, Ćirković-Veličković T. Tropomyosin quantification in seafood samples-right choice of standard makes a difference. in XXII EuroFoodChem conference, 14th-16th June, 2023. In: Book of Abstracts. 2023;:132-132.
https://hdl.handle.net/21.15107/rcub_cherry_6024 .

Krstić-Ristivojević, Maja, Jovanović, Vesna, Radomirović, Mirjana Ž., Trifunović, Olga, Stanić-Vučinić, Dragana, Ćirković-Veličković, Tanja, "Tropomyosin quantification in seafood samples-right choice of standard makes a difference" in XXII EuroFoodChem conference, 14th-16th June, 2023. In: Book of Abstracts (2023):132-132,
https://hdl.handle.net/21.15107/rcub_cherry_6024 .

TY  - JOUR
AU  - Veličković, Luka
AU  - Simović, Ana
AU  - Gligorijević, Nikola
AU  - Thureau, Aurélien
AU  - Obradović, Milica
AU  - Vasović, Tamara
AU  - Sotiroudis, Georgios
AU  - Zoumpanioti, Maria
AU  - Brûlet, Annie
AU  - Ćirković-Veličković, Tanja
AU  - Combet, Sophie
AU  - Nikolić, Milan
AU  - Minić, Simeon
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6273
AB  - This study aimed to purify, characterise and stabilise the natural food colourant, R-phycocyanin (R-PC), from the red algae Porphyra spp. (Nori). We purified R-PC from dried Nori flakes with a high purity ratio (A618/A280 ≥ 3.4) in native form (α-helix content 53%). SAXS measurements revealed that R-PC is trimeric ((αβ)3) in solution. The thermal denaturation of α-helix revealed one transition (Tm at 52 °C), while the pH stability study showed R-PC is stable in the pH range 4–8. The thermal treatment of R-PC at 60 °C has detrimental and irreversible effects on R-PC colour and antioxidant capacity (22 % of residual capacity). However, immobilisation of R-PC within calcium alginate beads completely preserves R-PC colour and mainly retains its antioxidant ability (78 % of residual capacity). Results give new insights into the stability of R-PC and preservation of its purple colour and bioactivity by encapsulation in calcium alginate beads.
PB  - Elsevier
T2  - Food Chemistry
T1  - Exploring and strengthening the potential of R-phycocyanin from Nori flakes as a food colourant
VL  - 426
SP  - 136669
DO  - 10.1016/j.foodchem.2023.136669
ER  - 

@article{
author = "Veličković, Luka and Simović, Ana and Gligorijević, Nikola and Thureau, Aurélien and Obradović, Milica and Vasović, Tamara and Sotiroudis, Georgios and Zoumpanioti, Maria and Brûlet, Annie and Ćirković-Veličković, Tanja and Combet, Sophie and Nikolić, Milan and Minić, Simeon",
year = "2023",
abstract = "This study aimed to purify, characterise and stabilise the natural food colourant, R-phycocyanin (R-PC), from the red algae Porphyra spp. (Nori). We purified R-PC from dried Nori flakes with a high purity ratio (A618/A280 ≥ 3.4) in native form (α-helix content 53%). SAXS measurements revealed that R-PC is trimeric ((αβ)3) in solution. The thermal denaturation of α-helix revealed one transition (Tm at 52 °C), while the pH stability study showed R-PC is stable in the pH range 4–8. The thermal treatment of R-PC at 60 °C has detrimental and irreversible effects on R-PC colour and antioxidant capacity (22 % of residual capacity). However, immobilisation of R-PC within calcium alginate beads completely preserves R-PC colour and mainly retains its antioxidant ability (78 % of residual capacity). Results give new insights into the stability of R-PC and preservation of its purple colour and bioactivity by encapsulation in calcium alginate beads.",
publisher = "Elsevier",
journal = "Food Chemistry",
title = "Exploring and strengthening the potential of R-phycocyanin from Nori flakes as a food colourant",
volume = "426",
pages = "136669",
doi = "10.1016/j.foodchem.2023.136669"
}

Veličković, L., Simović, A., Gligorijević, N., Thureau, A., Obradović, M., Vasović, T., Sotiroudis, G., Zoumpanioti, M., Brûlet, A., Ćirković-Veličković, T., Combet, S., Nikolić, M.,& Minić, S.. (2023). Exploring and strengthening the potential of R-phycocyanin from Nori flakes as a food colourant. in Food Chemistry
Elsevier., 426, 136669.
https://doi.org/10.1016/j.foodchem.2023.136669

Veličković L, Simović A, Gligorijević N, Thureau A, Obradović M, Vasović T, Sotiroudis G, Zoumpanioti M, Brûlet A, Ćirković-Veličković T, Combet S, Nikolić M, Minić S. Exploring and strengthening the potential of R-phycocyanin from Nori flakes as a food colourant. in Food Chemistry. 2023;426:136669.
doi:10.1016/j.foodchem.2023.136669 .

Veličković, Luka, Simović, Ana, Gligorijević, Nikola, Thureau, Aurélien, Obradović, Milica, Vasović, Tamara, Sotiroudis, Georgios, Zoumpanioti, Maria, Brûlet, Annie, Ćirković-Veličković, Tanja, Combet, Sophie, Nikolić, Milan, Minić, Simeon, "Exploring and strengthening the potential of R-phycocyanin from Nori flakes as a food colourant" in Food Chemistry, 426 (2023):136669,
https://doi.org/10.1016/j.foodchem.2023.136669 . .

TY  - CONF
AU  - Lujić, Tamara
AU  - Gligorijević, Nikola
AU  - Jovanović, Vesna
AU  - Aćimović, Jelena
AU  - Mitić, Dragana
AU  - Vasović, Tamara
AU  - Stojadinović, Marija
AU  - Stanić-Vučinić, Dragana
AU  - Ćirković-Veličković, Tanja
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6424
AB  - Microplastics is abundant in the environment, food and beverages and get ingested by humans. Its complex interplay with proteins lead to formation of corona. Tightly bound proteins represent hard corona, while weaker binding partners are found in soft corona. Separation of hard and soft corona of allergenic proteins of shrimps, eggs and cow’s milk, tropomyosin (TPM), ovalbumin (OVA) and beta-lactoglobulin (BLG) and identification of binding partners by proteomics was aim of our study.
Allergenic proteins were purified from egg white, shrimps and cow’s milk. Binding to polyethylene terephthalate microplastics (PET) (70-100 m) was probed at pH 7 for purified allergens and egg white proteins. After establishment of binding equilibrium, soft and hard corona were separated and analyzed by SDS PAGE, followed by identification of bound proteins by nanoLC-HRMS. Binding of all allergenic proteins was observed in both soft and hard corona. Soft corona contains exclusively intact, full length OVA, TPM and BLG. Hard corona is enriched for truncated OVA and oligomers of TPM. OVA fragments are partially or fully enfolded and have higher level of exposed hydrophobic patches resulting in higher affinity for PET microplastics. In comparison to OVA and TPM, hard corona of BLG is less abundant under similar conditions. BLG is compact globular protein with lower level of exposed hydrophobic patches in comparison to ovalbumin and tropomyosin. In hard corona, trace amounts of contaminating alfa-lactalbumin become enriched. In the presence of egg white protein extract OVA forms both SC and HC on microplastics, being the dominant protein of hard corona (with ovotransferrin). Lysozyme and ovomucin are present only in hard corona. Both proteins are known for their strong bioactivity and represent a small fraction of total egg white proteins.
Our results show that allergenic proteins form hard corona on PET microplastics. Among egg white proteins, minor proteins such as lysozyme and ovomucin become enriched. Denaturing effect of strong binding to microplastics may change functional characteristics of allergens and bioactive proteins of foods and should be further investigated in functional assays.
Acknowledgment: This study was supported by IMPTOX European Union's Horizon 2020 research and innovation program (grant number 965173).
PB  - Italian Proteomics Association
C3  - ItPA HPS and SePA XVII International Congress: Proteomics and Metabolomics towards Global Health, November 29th – December 1st, 2023, Roma, Italy
T1  - Proteomic insight into allergenic food corona on polyethylene terephthalate microplastics
SP  - 11
EP  - 11
UR  - https://hdl.handle.net/21.15107/rcub_cherry_6424
ER  - 

@conference{
author = "Lujić, Tamara and Gligorijević, Nikola and Jovanović, Vesna and Aćimović, Jelena and Mitić, Dragana and Vasović, Tamara and Stojadinović, Marija and Stanić-Vučinić, Dragana and Ćirković-Veličković, Tanja",
year = "2023",
abstract = "Microplastics is abundant in the environment, food and beverages and get ingested by humans. Its complex interplay with proteins lead to formation of corona. Tightly bound proteins represent hard corona, while weaker binding partners are found in soft corona. Separation of hard and soft corona of allergenic proteins of shrimps, eggs and cow’s milk, tropomyosin (TPM), ovalbumin (OVA) and beta-lactoglobulin (BLG) and identification of binding partners by proteomics was aim of our study.
Allergenic proteins were purified from egg white, shrimps and cow’s milk. Binding to polyethylene terephthalate microplastics (PET) (70-100 m) was probed at pH 7 for purified allergens and egg white proteins. After establishment of binding equilibrium, soft and hard corona were separated and analyzed by SDS PAGE, followed by identification of bound proteins by nanoLC-HRMS. Binding of all allergenic proteins was observed in both soft and hard corona. Soft corona contains exclusively intact, full length OVA, TPM and BLG. Hard corona is enriched for truncated OVA and oligomers of TPM. OVA fragments are partially or fully enfolded and have higher level of exposed hydrophobic patches resulting in higher affinity for PET microplastics. In comparison to OVA and TPM, hard corona of BLG is less abundant under similar conditions. BLG is compact globular protein with lower level of exposed hydrophobic patches in comparison to ovalbumin and tropomyosin. In hard corona, trace amounts of contaminating alfa-lactalbumin become enriched. In the presence of egg white protein extract OVA forms both SC and HC on microplastics, being the dominant protein of hard corona (with ovotransferrin). Lysozyme and ovomucin are present only in hard corona. Both proteins are known for their strong bioactivity and represent a small fraction of total egg white proteins.
Our results show that allergenic proteins form hard corona on PET microplastics. Among egg white proteins, minor proteins such as lysozyme and ovomucin become enriched. Denaturing effect of strong binding to microplastics may change functional characteristics of allergens and bioactive proteins of foods and should be further investigated in functional assays.
Acknowledgment: This study was supported by IMPTOX European Union's Horizon 2020 research and innovation program (grant number 965173).",
publisher = "Italian Proteomics Association",
journal = "ItPA HPS and SePA XVII International Congress: Proteomics and Metabolomics towards Global Health, November 29th – December 1st, 2023, Roma, Italy",
title = "Proteomic insight into allergenic food corona on polyethylene terephthalate microplastics",
pages = "11-11",
url = "https://hdl.handle.net/21.15107/rcub_cherry_6424"
}

Lujić, T., Gligorijević, N., Jovanović, V., Aćimović, J., Mitić, D., Vasović, T., Stojadinović, M., Stanić-Vučinić, D.,& Ćirković-Veličković, T.. (2023). Proteomic insight into allergenic food corona on polyethylene terephthalate microplastics. in ItPA HPS and SePA XVII International Congress: Proteomics and Metabolomics towards Global Health, November 29th – December 1st, 2023, Roma, Italy
Italian Proteomics Association., 11-11.
https://hdl.handle.net/21.15107/rcub_cherry_6424

Lujić T, Gligorijević N, Jovanović V, Aćimović J, Mitić D, Vasović T, Stojadinović M, Stanić-Vučinić D, Ćirković-Veličković T. Proteomic insight into allergenic food corona on polyethylene terephthalate microplastics. in ItPA HPS and SePA XVII International Congress: Proteomics and Metabolomics towards Global Health, November 29th – December 1st, 2023, Roma, Italy. 2023;:11-11.
https://hdl.handle.net/21.15107/rcub_cherry_6424 .

Lujić, Tamara, Gligorijević, Nikola, Jovanović, Vesna, Aćimović, Jelena, Mitić, Dragana, Vasović, Tamara, Stojadinović, Marija, Stanić-Vučinić, Dragana, Ćirković-Veličković, Tanja, "Proteomic insight into allergenic food corona on polyethylene terephthalate microplastics" in ItPA HPS and SePA XVII International Congress: Proteomics and Metabolomics towards Global Health, November 29th – December 1st, 2023, Roma, Italy (2023):11-11,
https://hdl.handle.net/21.15107/rcub_cherry_6424 .

TY  - CONF
AU  - Simović, Ana
AU  - Bićanin, Maša
AU  - Radomirović, Mirjana Ž.
AU  - Aćimovič, Jelena
AU  - Mitić, Dragana
AU  - Stanić-Vučinić, Dragana
AU  - Ćirković-Veličković, Tanja
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6418
AB  - The emergence of the novel coronavirus SARS-CoV-2 has attracted the attention of the whole scientific community. However, as there are significant concerns regarding the effectiveness of vaccines and drugs against novel SARS-CoV-2 variants, naturally derived broad-spectrum of antivirals seems to be precious adjuvant agents to assist in combat against this disease. Phycocyanobilin (PCB) is an open-chain tetrapyrrole chromophore of phycocyanin (PC), chromoprotein derived from Spirullina, with strong anti-oxidative action. The role of disulfide bonds and thiol-disulfide balance in RBD is considered to play a significant role in the binding of S protein to ACE2 receptor. In RBD, in contrast to C480–C488 disulfide, which is thermodynamically stable, C379–C432 and C391–C525 disulfides are in dynamic equilibrium with their thiol states and, thus these two pairs of disulfides are more sensitive to changes in redox poise. Our study aimed to investigate impact of PCB on disulfide balance of RBD by redox proteomics and to investigate structural changes in the protein exposed to PCB. The effect of PCB on RBD secondary structures was examined by far-UV CD spectroscopy after titration of RBD with increasing concentrations of PCB. The presence of PCB had a pronounced effect on the spectral shape. RBD is dominantly composed of random coils and β-sheets. In the presence of PCB a slight increase of α-helical and random coils content, while the content of β-sheets and β-turns is decreased. Mapping redox-active disulfides and reactive cysteines in recombinant SARS-CoV-2 RBD was done using redox proteomics on both recombinant RBD and PCB-exposed RBD. A mass shift caused by alkylation of free Cys residues was detected on three Cys residues demonstrating disulfides C379–C432 and 432-391 to be semi-stable in both RBD and PCB-exposed RBD. Our results demonstrate that RBD exposed to PCB undergo structural changes but does not change the redox state of its critical semi-stable disulfides.
Acknowledgment: The authors acknowledge support of the special Research Fund (BOF) of Ghent University (grant number 01N01718) and the Ministry of Science, Innovation and technological development of the Republic of Serbia (Contract number: 451-03-68/2023-14/200168).
PB  - Italian Proteomics Association
C3  - ItPA HPS and SePA XVII International Congress: Proteomics and Metabolomics towards Global Health, November 29th – December 1st, 2023, Roma, Italy
T1  - Redox status of critical disulfides of SARS-CoV-2 receptor-binding-domain exposed to bioactive chomophore phycocyanobilin
SP  - 63
EP  - 63
UR  - https://hdl.handle.net/21.15107/rcub_cherry_6418
ER  - 

@conference{
author = "Simović, Ana and Bićanin, Maša and Radomirović, Mirjana Ž. and Aćimovič, Jelena and Mitić, Dragana and Stanić-Vučinić, Dragana and Ćirković-Veličković, Tanja",
year = "2023",
abstract = "The emergence of the novel coronavirus SARS-CoV-2 has attracted the attention of the whole scientific community. However, as there are significant concerns regarding the effectiveness of vaccines and drugs against novel SARS-CoV-2 variants, naturally derived broad-spectrum of antivirals seems to be precious adjuvant agents to assist in combat against this disease. Phycocyanobilin (PCB) is an open-chain tetrapyrrole chromophore of phycocyanin (PC), chromoprotein derived from Spirullina, with strong anti-oxidative action. The role of disulfide bonds and thiol-disulfide balance in RBD is considered to play a significant role in the binding of S protein to ACE2 receptor. In RBD, in contrast to C480–C488 disulfide, which is thermodynamically stable, C379–C432 and C391–C525 disulfides are in dynamic equilibrium with their thiol states and, thus these two pairs of disulfides are more sensitive to changes in redox poise. Our study aimed to investigate impact of PCB on disulfide balance of RBD by redox proteomics and to investigate structural changes in the protein exposed to PCB. The effect of PCB on RBD secondary structures was examined by far-UV CD spectroscopy after titration of RBD with increasing concentrations of PCB. The presence of PCB had a pronounced effect on the spectral shape. RBD is dominantly composed of random coils and β-sheets. In the presence of PCB a slight increase of α-helical and random coils content, while the content of β-sheets and β-turns is decreased. Mapping redox-active disulfides and reactive cysteines in recombinant SARS-CoV-2 RBD was done using redox proteomics on both recombinant RBD and PCB-exposed RBD. A mass shift caused by alkylation of free Cys residues was detected on three Cys residues demonstrating disulfides C379–C432 and 432-391 to be semi-stable in both RBD and PCB-exposed RBD. Our results demonstrate that RBD exposed to PCB undergo structural changes but does not change the redox state of its critical semi-stable disulfides.
Acknowledgment: The authors acknowledge support of the special Research Fund (BOF) of Ghent University (grant number 01N01718) and the Ministry of Science, Innovation and technological development of the Republic of Serbia (Contract number: 451-03-68/2023-14/200168).",
publisher = "Italian Proteomics Association",
journal = "ItPA HPS and SePA XVII International Congress: Proteomics and Metabolomics towards Global Health, November 29th – December 1st, 2023, Roma, Italy",
title = "Redox status of critical disulfides of SARS-CoV-2 receptor-binding-domain exposed to bioactive chomophore phycocyanobilin",
pages = "63-63",
url = "https://hdl.handle.net/21.15107/rcub_cherry_6418"
}

Simović, A., Bićanin, M., Radomirović, M. Ž., Aćimovič, J., Mitić, D., Stanić-Vučinić, D.,& Ćirković-Veličković, T.. (2023). Redox status of critical disulfides of SARS-CoV-2 receptor-binding-domain exposed to bioactive chomophore phycocyanobilin. in ItPA HPS and SePA XVII International Congress: Proteomics and Metabolomics towards Global Health, November 29th – December 1st, 2023, Roma, Italy
Italian Proteomics Association., 63-63.
https://hdl.handle.net/21.15107/rcub_cherry_6418

Simović A, Bićanin M, Radomirović MŽ, Aćimovič J, Mitić D, Stanić-Vučinić D, Ćirković-Veličković T. Redox status of critical disulfides of SARS-CoV-2 receptor-binding-domain exposed to bioactive chomophore phycocyanobilin. in ItPA HPS and SePA XVII International Congress: Proteomics and Metabolomics towards Global Health, November 29th – December 1st, 2023, Roma, Italy. 2023;:63-63.
https://hdl.handle.net/21.15107/rcub_cherry_6418 .

Simović, Ana, Bićanin, Maša, Radomirović, Mirjana Ž., Aćimovič, Jelena, Mitić, Dragana, Stanić-Vučinić, Dragana, Ćirković-Veličković, Tanja, "Redox status of critical disulfides of SARS-CoV-2 receptor-binding-domain exposed to bioactive chomophore phycocyanobilin" in ItPA HPS and SePA XVII International Congress: Proteomics and Metabolomics towards Global Health, November 29th – December 1st, 2023, Roma, Italy (2023):63-63,
https://hdl.handle.net/21.15107/rcub_cherry_6418 .

TY  - JOUR
AU  - Veličković, Luka
AU  - Simović, Ana
AU  - Gligorijević, Nikola
AU  - Thureau, Aurelien
AU  - Obradović, Milica
AU  - Vasović, Tamara
AU  - Sotiroudis, Georgios
AU  - Zoumpanioti, Maria
AU  - Brûlet, Annie
AU  - Ćirković-Veličković, Tanja
AU  - Combet, Sophie
AU  - Nikolić, Milan
AU  - Minić, Simeon
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6420
AB  - This study aimed to purify, characterise and stabilise the natural food colourant, R-phycocyanin (R-PC), from thered algae Porphyra spp. (Nori). We purified R-PC from dried Nori flakes with a high purity ratio (A618/A280 ≥ 3.4)in native form (α-helix content 53%). SAXS measurements revealed that R-PC is trimeric ((αβ)3) in solution. Thethermal denaturation of α-helix revealed one transition (Tm at 52 ◦C), while the pH stability study showed R-PC isstable in the pH range 4–8. The thermal treatment of R-PC at 60 ◦C has detrimental and irreversible effects on RPCcolour and antioxidant capacity (22 % of residual capacity). However, immobilisation of R-PC within calciumalginate beads completely preserves R-PC colour and mainly retains its antioxidant ability (78 % of residualcapacity). Results give new insights into the stability of R-PC and preservation of its purple colour and bioactivityby encapsulation in calcium alginate beads.
PB  - Elsevier
T2  - Food Chemistry
T1  - Exploring and strengthening the potential of R-phycocyanin from Nori flakes as a food colourant
VL  - 426
SP  - 136669
DO  - 10.1016/j.foodchem.2023.136669
ER  - 

@article{
author = "Veličković, Luka and Simović, Ana and Gligorijević, Nikola and Thureau, Aurelien and Obradović, Milica and Vasović, Tamara and Sotiroudis, Georgios and Zoumpanioti, Maria and Brûlet, Annie and Ćirković-Veličković, Tanja and Combet, Sophie and Nikolić, Milan and Minić, Simeon",
year = "2023",
abstract = "This study aimed to purify, characterise and stabilise the natural food colourant, R-phycocyanin (R-PC), from thered algae Porphyra spp. (Nori). We purified R-PC from dried Nori flakes with a high purity ratio (A618/A280 ≥ 3.4)in native form (α-helix content 53%). SAXS measurements revealed that R-PC is trimeric ((αβ)3) in solution. Thethermal denaturation of α-helix revealed one transition (Tm at 52 ◦C), while the pH stability study showed R-PC isstable in the pH range 4–8. The thermal treatment of R-PC at 60 ◦C has detrimental and irreversible effects on RPCcolour and antioxidant capacity (22 % of residual capacity). However, immobilisation of R-PC within calciumalginate beads completely preserves R-PC colour and mainly retains its antioxidant ability (78 % of residualcapacity). Results give new insights into the stability of R-PC and preservation of its purple colour and bioactivityby encapsulation in calcium alginate beads.",
publisher = "Elsevier",
journal = "Food Chemistry",
title = "Exploring and strengthening the potential of R-phycocyanin from Nori flakes as a food colourant",
volume = "426",
pages = "136669",
doi = "10.1016/j.foodchem.2023.136669"
}

Veličković, L., Simović, A., Gligorijević, N., Thureau, A., Obradović, M., Vasović, T., Sotiroudis, G., Zoumpanioti, M., Brûlet, A., Ćirković-Veličković, T., Combet, S., Nikolić, M.,& Minić, S.. (2023). Exploring and strengthening the potential of R-phycocyanin from Nori flakes as a food colourant. in Food Chemistry
Elsevier., 426, 136669.
https://doi.org/10.1016/j.foodchem.2023.136669

Veličković L, Simović A, Gligorijević N, Thureau A, Obradović M, Vasović T, Sotiroudis G, Zoumpanioti M, Brûlet A, Ćirković-Veličković T, Combet S, Nikolić M, Minić S. Exploring and strengthening the potential of R-phycocyanin from Nori flakes as a food colourant. in Food Chemistry. 2023;426:136669.
doi:10.1016/j.foodchem.2023.136669 .

Veličković, Luka, Simović, Ana, Gligorijević, Nikola, Thureau, Aurelien, Obradović, Milica, Vasović, Tamara, Sotiroudis, Georgios, Zoumpanioti, Maria, Brûlet, Annie, Ćirković-Veličković, Tanja, Combet, Sophie, Nikolić, Milan, Minić, Simeon, "Exploring and strengthening the potential of R-phycocyanin from Nori flakes as a food colourant" in Food Chemistry, 426 (2023):136669,
https://doi.org/10.1016/j.foodchem.2023.136669 . .

TY  - JOUR
AU  - Mladenović Stokanić, Maja
AU  - Simović, Ana
AU  - Jovanović, Vesna B.
AU  - Radomirović, Mirjana
AU  - Udovićki, Božidar
AU  - Krstić Ristivojević, Maja
AU  - Đukić, Teodora
AU  - Vasović, Tamara
AU  - Aćimović, Jelena
AU  - Sabljić, Ljiljana
AU  - Lukić, Ivana
AU  - Kovačević, Ana
AU  - Cujić, Danica
AU  - Gnjatović, Marija
AU  - Smiljanić, Katarina
AU  - Stojadinović, Marija
AU  - Radosavljević, Jelena
AU  - Stanić-Vučinić, Dragana
AU  - Stojanović, Marijana
AU  - Rajković, Andreja
AU  - Ćirković-Veličković, Tanja
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6436
AB  - In this study, a cost-effective sandwich ELISA test, based on polyclonal antibodies, for routine quantification SARS-CoV-2 nucleocapsid (N) protein was developed. The recombinant N protein was produced and used for the production of mice and rabbit antisera. Polyclonal N protein-specific antibodies served as capture and detection antibodies. The prototype ELISA has LOD 0.93 ng/mL and LOQ 5.3 ng/mL, with a linear range of 1.52–48.83 ng/mL. N protein heat pretreatment (56 °C, 1 h) decreased, while pretreatment with 1% Triton X-100 increased analytical ELISA sensitivity. The diagnostic specificity of ELISA was 100% (95% CI, 91.19–100.00%) and sensitivity was 52.94% (95% CI, 35.13–70.22%) compared to rtRT-PCR (Ct < 40). Profoundly higher sensitivity was obtained using patient samples mostly containing Wuhan-similar variants (Wuhan, alpha, and delta), 62.50% (95% CI, 40.59 to 81.20%), in comparison to samples mostly containing Wuhan-distant variants (Omicron) 30.00% (6.67–65.25%). The developed product has relatively high diagnostic sensitivity in relation to its analytical sensitivity due to the usage of polyclonal antibodies from two species, providing a wide repertoire of antibodies against multiple N protein epitopes. Moreover, the fast, simple, and inexpensive production of polyclonal antibodies, as the most expensive assay components, would result in affordable antigen tests.
PB  - MDPI
T2  - International Journal of Molecular Sciences
T1  - Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species
VL  - 25
IS  - 1
SP  - 333
DO  - 10.3390/ijms25010333
ER  - 

@article{
author = "Mladenović Stokanić, Maja and Simović, Ana and Jovanović, Vesna B. and Radomirović, Mirjana and Udovićki, Božidar and Krstić Ristivojević, Maja and Đukić, Teodora and Vasović, Tamara and Aćimović, Jelena and Sabljić, Ljiljana and Lukić, Ivana and Kovačević, Ana and Cujić, Danica and Gnjatović, Marija and Smiljanić, Katarina and Stojadinović, Marija and Radosavljević, Jelena and Stanić-Vučinić, Dragana and Stojanović, Marijana and Rajković, Andreja and Ćirković-Veličković, Tanja",
year = "2023",
abstract = "In this study, a cost-effective sandwich ELISA test, based on polyclonal antibodies, for routine quantification SARS-CoV-2 nucleocapsid (N) protein was developed. The recombinant N protein was produced and used for the production of mice and rabbit antisera. Polyclonal N protein-specific antibodies served as capture and detection antibodies. The prototype ELISA has LOD 0.93 ng/mL and LOQ 5.3 ng/mL, with a linear range of 1.52–48.83 ng/mL. N protein heat pretreatment (56 °C, 1 h) decreased, while pretreatment with 1% Triton X-100 increased analytical ELISA sensitivity. The diagnostic specificity of ELISA was 100% (95% CI, 91.19–100.00%) and sensitivity was 52.94% (95% CI, 35.13–70.22%) compared to rtRT-PCR (Ct < 40). Profoundly higher sensitivity was obtained using patient samples mostly containing Wuhan-similar variants (Wuhan, alpha, and delta), 62.50% (95% CI, 40.59 to 81.20%), in comparison to samples mostly containing Wuhan-distant variants (Omicron) 30.00% (6.67–65.25%). The developed product has relatively high diagnostic sensitivity in relation to its analytical sensitivity due to the usage of polyclonal antibodies from two species, providing a wide repertoire of antibodies against multiple N protein epitopes. Moreover, the fast, simple, and inexpensive production of polyclonal antibodies, as the most expensive assay components, would result in affordable antigen tests.",
publisher = "MDPI",
journal = "International Journal of Molecular Sciences",
title = "Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species",
volume = "25",
number = "1",
pages = "333",
doi = "10.3390/ijms25010333"
}

Mladenović Stokanić, M., Simović, A., Jovanović, V. B., Radomirović, M., Udovićki, B., Krstić Ristivojević, M., Đukić, T., Vasović, T., Aćimović, J., Sabljić, L., Lukić, I., Kovačević, A., Cujić, D., Gnjatović, M., Smiljanić, K., Stojadinović, M., Radosavljević, J., Stanić-Vučinić, D., Stojanović, M., Rajković, A.,& Ćirković-Veličković, T.. (2023). Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species. in International Journal of Molecular Sciences
MDPI., 25(1), 333.
https://doi.org/10.3390/ijms25010333

Mladenović Stokanić M, Simović A, Jovanović VB, Radomirović M, Udovićki B, Krstić Ristivojević M, Đukić T, Vasović T, Aćimović J, Sabljić L, Lukić I, Kovačević A, Cujić D, Gnjatović M, Smiljanić K, Stojadinović M, Radosavljević J, Stanić-Vučinić D, Stojanović M, Rajković A, Ćirković-Veličković T. Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species. in International Journal of Molecular Sciences. 2023;25(1):333.
doi:10.3390/ijms25010333 .

Mladenović Stokanić, Maja, Simović, Ana, Jovanović, Vesna B., Radomirović, Mirjana, Udovićki, Božidar, Krstić Ristivojević, Maja, Đukić, Teodora, Vasović, Tamara, Aćimović, Jelena, Sabljić, Ljiljana, Lukić, Ivana, Kovačević, Ana, Cujić, Danica, Gnjatović, Marija, Smiljanić, Katarina, Stojadinović, Marija, Radosavljević, Jelena, Stanić-Vučinić, Dragana, Stojanović, Marijana, Rajković, Andreja, Ćirković-Veličković, Tanja, "Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species" in International Journal of Molecular Sciences, 25, no. 1 (2023):333,
https://doi.org/10.3390/ijms25010333 . .

TY  - DATA
AU  - Radomirović, Mirjana
AU  - Gligorijević, Nikola
AU  - Stanić-Vučinić, Dragana
AU  - Rajković, Andreja
AU  - Ćirković-Veličković, Tanja
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6466
AB  - Experimental data for the results shown in the manuscript Ultrasensitive Quantification of Crustacean Tropomyosin by Immuno-PCR are given in the attachment.
PB  - MDPI
T2  - International Journal of Molecular Sciences
T1  - Research data for: Radomirović, M. Ž., Gligorijević, N., Stanić-Vučinić, D., Rajković, A.,& Ćirković-Veličković, T.. (2023). Ultrasensitive Quantification of Crustacean Tropomyosin by Immuno-PCR. in International Journal of Molecular Sciences MDPI., 24(20), 15410. https://doi.org/doi.org/10.3390/ ijms242015410
VL  - 24
IS  - 20
UR  - https://hdl.handle.net/21.15107/rcub_cherry_6466
ER  - 

@misc{
author = "Radomirović, Mirjana and Gligorijević, Nikola and Stanić-Vučinić, Dragana and Rajković, Andreja and Ćirković-Veličković, Tanja",
year = "2023",
abstract = "Experimental data for the results shown in the manuscript Ultrasensitive Quantification of Crustacean Tropomyosin by Immuno-PCR are given in the attachment.",
publisher = "MDPI",
journal = "International Journal of Molecular Sciences",
title = "Research data for: Radomirović, M. Ž., Gligorijević, N., Stanić-Vučinić, D., Rajković, A.,& Ćirković-Veličković, T.. (2023). Ultrasensitive Quantification of Crustacean Tropomyosin by Immuno-PCR. in International Journal of Molecular Sciences MDPI., 24(20), 15410. https://doi.org/doi.org/10.3390/ ijms242015410",
volume = "24",
number = "20",
url = "https://hdl.handle.net/21.15107/rcub_cherry_6466"
}

Radomirović, M., Gligorijević, N., Stanić-Vučinić, D., Rajković, A.,& Ćirković-Veličković, T.. (2023). Research data for: Radomirović, M. Ž., Gligorijević, N., Stanić-Vučinić, D., Rajković, A.,& Ćirković-Veličković, T.. (2023). Ultrasensitive Quantification of Crustacean Tropomyosin by Immuno-PCR. in International Journal of Molecular Sciences MDPI., 24(20), 15410. https://doi.org/doi.org/10.3390/ ijms242015410. in International Journal of Molecular Sciences
MDPI., 24(20).
https://hdl.handle.net/21.15107/rcub_cherry_6466

Radomirović M, Gligorijević N, Stanić-Vučinić D, Rajković A, Ćirković-Veličković T. Research data for: Radomirović, M. Ž., Gligorijević, N., Stanić-Vučinić, D., Rajković, A.,& Ćirković-Veličković, T.. (2023). Ultrasensitive Quantification of Crustacean Tropomyosin by Immuno-PCR. in International Journal of Molecular Sciences MDPI., 24(20), 15410. https://doi.org/doi.org/10.3390/ ijms242015410. in International Journal of Molecular Sciences. 2023;24(20).
https://hdl.handle.net/21.15107/rcub_cherry_6466 .

Radomirović, Mirjana, Gligorijević, Nikola, Stanić-Vučinić, Dragana, Rajković, Andreja, Ćirković-Veličković, Tanja, "Research data for: Radomirović, M. Ž., Gligorijević, N., Stanić-Vučinić, D., Rajković, A.,& Ćirković-Veličković, T.. (2023). Ultrasensitive Quantification of Crustacean Tropomyosin by Immuno-PCR. in International Journal of Molecular Sciences MDPI., 24(20), 15410. https://doi.org/doi.org/10.3390/ ijms242015410" in International Journal of Molecular Sciences, 24, no. 20 (2023),
https://hdl.handle.net/21.15107/rcub_cherry_6466 .

TY  - JOUR
AU  - Radomirović, Mirjana Ž.
AU  - Gligorijević, Nikola
AU  - Stanić-Vučinić, Dragana
AU  - Rajković, Andreja
AU  - Ćirković-Veličković, Tanja
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6019
AB  - Tropomyosin is the major and predominant allergen among shellfish. This study developed
an ultrasensitive immuno-PCR method for the quantification of crustacean tropomyosin in foods.
The method couples sandwich ELISA with the real-time PCR (rtPCR) amplification of marker DNAs. Monoclonal anti-TPM antibody was the capture antibody, polyclonal rabbit anti-shrimp tropomyosin antibody was the detection antibody, while natural shrimp tropomyosin served as the standard. A double-stranded amino-DNA was covalently conjugated to a secondary anti-rabbit antibody and subsequently amplified and quantified via rtPCR. The quantification sensitivity of immuno-PCR was 20-fold higher than analogous ELISA, with LOQ 19.8 pg/mL. The developed immuno-PCR method is highly specific for the detection of crustacean tropomyosin and is highly precise in a broad concentration range. Tropomyosin recovery in the spiked vegetable soup was 87.7–115.6%. Crustacean tropomyosin was also quantified in commercial food products. The reported immuno-PCR assay is the most sensitive method for the quantification of crustacean tropomyosin and is the first immuno-PCR-based assay for the quantification of food allergen and food protein in general. The described method could be easily adapted for the specific and ultrasensitive immuno-PCR-based detection of traces of any food allergen that is currently being quantified with ELISA, which is of critical importance for people with food allergies.
PB  - MDPI
T2  - International Journal of Molecular Sciences
T1  - Ultrasensitive Quantification of Crustacean Tropomyosin by Immuno-PCR
VL  - 24
IS  - 20
SP  - 15410
DO  - doi.org/10.3390/ ijms242015410
ER  - 

@article{
author = "Radomirović, Mirjana Ž. and Gligorijević, Nikola and Stanić-Vučinić, Dragana and Rajković, Andreja and Ćirković-Veličković, Tanja",
year = "2023",
abstract = "Tropomyosin is the major and predominant allergen among shellfish. This study developed
an ultrasensitive immuno-PCR method for the quantification of crustacean tropomyosin in foods.
The method couples sandwich ELISA with the real-time PCR (rtPCR) amplification of marker DNAs. Monoclonal anti-TPM antibody was the capture antibody, polyclonal rabbit anti-shrimp tropomyosin antibody was the detection antibody, while natural shrimp tropomyosin served as the standard. A double-stranded amino-DNA was covalently conjugated to a secondary anti-rabbit antibody and subsequently amplified and quantified via rtPCR. The quantification sensitivity of immuno-PCR was 20-fold higher than analogous ELISA, with LOQ 19.8 pg/mL. The developed immuno-PCR method is highly specific for the detection of crustacean tropomyosin and is highly precise in a broad concentration range. Tropomyosin recovery in the spiked vegetable soup was 87.7–115.6%. Crustacean tropomyosin was also quantified in commercial food products. The reported immuno-PCR assay is the most sensitive method for the quantification of crustacean tropomyosin and is the first immuno-PCR-based assay for the quantification of food allergen and food protein in general. The described method could be easily adapted for the specific and ultrasensitive immuno-PCR-based detection of traces of any food allergen that is currently being quantified with ELISA, which is of critical importance for people with food allergies.",
publisher = "MDPI",
journal = "International Journal of Molecular Sciences",
title = "Ultrasensitive Quantification of Crustacean Tropomyosin by Immuno-PCR",
volume = "24",
number = "20",
pages = "15410",
doi = "doi.org/10.3390/ ijms242015410"
}

Radomirović, M. Ž., Gligorijević, N., Stanić-Vučinić, D., Rajković, A.,& Ćirković-Veličković, T.. (2023). Ultrasensitive Quantification of Crustacean Tropomyosin by Immuno-PCR. in International Journal of Molecular Sciences
MDPI., 24(20), 15410.
https://doi.org/doi.org/10.3390/ ijms242015410

Radomirović MŽ, Gligorijević N, Stanić-Vučinić D, Rajković A, Ćirković-Veličković T. Ultrasensitive Quantification of Crustacean Tropomyosin by Immuno-PCR. in International Journal of Molecular Sciences. 2023;24(20):15410.
doi:doi.org/10.3390/ ijms242015410 .

Radomirović, Mirjana Ž., Gligorijević, Nikola, Stanić-Vučinić, Dragana, Rajković, Andreja, Ćirković-Veličković, Tanja, "Ultrasensitive Quantification of Crustacean Tropomyosin by Immuno-PCR" in International Journal of Molecular Sciences, 24, no. 20 (2023):15410,
https://doi.org/doi.org/10.3390/ ijms242015410 . .

TY  - JOUR
AU  - Abdelhameed, Shorok A. M.
AU  - de Azambuja, Francisco
AU  - Vasović, Tamara
AU  - Savić, Nada D.
AU  - Ćirković-Veličković, Tanja
AU  - Parac-Vogt, Tatjana N.
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/5816
AB  - Oxidative modifications of proteins are key to many applications in biotechnology. Metal-catalyzed oxidation reactions efficiently oxidize proteins but with low selectivity, and are highly dependent on the protein surface residues to direct the reaction. Herein, we demonstrate that discrete inorganic ligands such as polyoxometalates enable an efficient and selective protein oxidative cleavage. In the presence of ascorbate (1 mM), the Cu-substituted polyoxometalate K8[Cu2+(H2O)(α2-P2W17O61)], (CuIIWD, 0.05 mM) selectively cleave hen egg white lysozyme under physiological conditions (pH =7.5, 37 °C) producing only four bands in the gel electropherogram (12.7, 11, 10, and 5 kDa). Liquid chromatography/mass spectrometry analysis reveals a regioselective cleavage in the vicinity of crystallographic CuIIWD/lysozyme interaction sites. Mechanistically, polyoxometalate is critical to position the Cu at the protein surface and limit the generation of oxidative species to the proximity of binding sites. Ultimately, this study outlines the potential of discrete, designable metal oxo clusters as catalysts for the selective modification of proteins through radical mechanisms under non-denaturing conditions.
PB  - Springer Nature
T2  - Nature Communications
T1  - Regioselective protein oxidative cleavage enabled by enzyme-like recognition of an inorganic metal oxo cluster ligand
VL  - 14
IS  - 1
SP  - 486
DO  - 10.1038/s41467-023-36085-z
ER  - 

@article{
author = "Abdelhameed, Shorok A. M. and de Azambuja, Francisco and Vasović, Tamara and Savić, Nada D. and Ćirković-Veličković, Tanja and Parac-Vogt, Tatjana N.",
year = "2023",
abstract = "Oxidative modifications of proteins are key to many applications in biotechnology. Metal-catalyzed oxidation reactions efficiently oxidize proteins but with low selectivity, and are highly dependent on the protein surface residues to direct the reaction. Herein, we demonstrate that discrete inorganic ligands such as polyoxometalates enable an efficient and selective protein oxidative cleavage. In the presence of ascorbate (1 mM), the Cu-substituted polyoxometalate K8[Cu2+(H2O)(α2-P2W17O61)], (CuIIWD, 0.05 mM) selectively cleave hen egg white lysozyme under physiological conditions (pH =7.5, 37 °C) producing only four bands in the gel electropherogram (12.7, 11, 10, and 5 kDa). Liquid chromatography/mass spectrometry analysis reveals a regioselective cleavage in the vicinity of crystallographic CuIIWD/lysozyme interaction sites. Mechanistically, polyoxometalate is critical to position the Cu at the protein surface and limit the generation of oxidative species to the proximity of binding sites. Ultimately, this study outlines the potential of discrete, designable metal oxo clusters as catalysts for the selective modification of proteins through radical mechanisms under non-denaturing conditions.",
publisher = "Springer Nature",
journal = "Nature Communications",
title = "Regioselective protein oxidative cleavage enabled by enzyme-like recognition of an inorganic metal oxo cluster ligand",
volume = "14",
number = "1",
pages = "486",
doi = "10.1038/s41467-023-36085-z"
}

Abdelhameed, S. A. M., de Azambuja, F., Vasović, T., Savić, N. D., Ćirković-Veličković, T.,& Parac-Vogt, T. N.. (2023). Regioselective protein oxidative cleavage enabled by enzyme-like recognition of an inorganic metal oxo cluster ligand. in Nature Communications
Springer Nature., 14(1), 486.
https://doi.org/10.1038/s41467-023-36085-z

Abdelhameed SAM, de Azambuja F, Vasović T, Savić ND, Ćirković-Veličković T, Parac-Vogt TN. Regioselective protein oxidative cleavage enabled by enzyme-like recognition of an inorganic metal oxo cluster ligand. in Nature Communications. 2023;14(1):486.
doi:10.1038/s41467-023-36085-z .

Abdelhameed, Shorok A. M., de Azambuja, Francisco, Vasović, Tamara, Savić, Nada D., Ćirković-Veličković, Tanja, Parac-Vogt, Tatjana N., "Regioselective protein oxidative cleavage enabled by enzyme-like recognition of an inorganic metal oxo cluster ligand" in Nature Communications, 14, no. 1 (2023):486,
https://doi.org/10.1038/s41467-023-36085-z . .

TY  - DATA
AU  - Abdelhameed, Shorok A. M.
AU  - de Azambuja, Francisco
AU  - Vasović, Tamara
AU  - Savić, Nada D.
AU  - Ćirković-Veličković, Tanja
AU  - Parac-Vogt, Tatjana N.
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/5816
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/5840
AB  - Oxidative modifications of proteins are key to many applications in biotechnology. Metal-catalyzed oxidation reactions efficiently oxidize proteins but with low selectivity, and are highly dependent on the protein surface residues to direct the reaction. Herein, we demonstrate that discrete inorganic ligands such as polyoxometalates enable an efficient and selective protein oxidative cleavage. In the presence of ascorbate (1 mM), the Cu-substituted polyoxometalate K8[Cu2+(H2O)(α2-P2W17O61)], (CuIIWD, 0.05 mM) selectively cleave hen egg white lysozyme under physiological conditions (pH =7.5, 37 °C) producing only four bands in the gel electropherogram (12.7, 11, 10, and 5 kDa). Liquid chromatography/mass spectrometry analysis reveals a regioselective cleavage in the vicinity of crystallographic CuIIWD/lysozyme interaction sites. Mechanistically, polyoxometalate is critical to position the Cu at the protein surface and limit the generation of oxidative species to the proximity of binding sites. Ultimately, this study outlines the potential of discrete, designable metal oxo clusters as catalysts for the selective modification of proteins through radical mechanisms under non-denaturing conditions.
PB  - Springer Nature
T2  - Nature Communications
T1  - Supplementary material for: Abdelhameed, S. A. M., de Azambuja, F., Vasović, T., Savić, N. D., Ćirković-Veličković, T.,& Parac-Vogt, T. N.. (2023). Regioselective protein oxidative cleavage enabled by enzyme-like recognition of an inorganic metal oxo cluster ligand. in Nature Communications Springer Nature., 14(1), 486. https://doi.org/10.1038/s41467-023-36085-z
VL  - 14
IS  - 1
SP  - 486
UR  - https://hdl.handle.net/21.15107/rcub_cherry_5840
ER  - 

@misc{
author = "Abdelhameed, Shorok A. M. and de Azambuja, Francisco and Vasović, Tamara and Savić, Nada D. and Ćirković-Veličković, Tanja and Parac-Vogt, Tatjana N.",
year = "2023",
abstract = "Oxidative modifications of proteins are key to many applications in biotechnology. Metal-catalyzed oxidation reactions efficiently oxidize proteins but with low selectivity, and are highly dependent on the protein surface residues to direct the reaction. Herein, we demonstrate that discrete inorganic ligands such as polyoxometalates enable an efficient and selective protein oxidative cleavage. In the presence of ascorbate (1 mM), the Cu-substituted polyoxometalate K8[Cu2+(H2O)(α2-P2W17O61)], (CuIIWD, 0.05 mM) selectively cleave hen egg white lysozyme under physiological conditions (pH =7.5, 37 °C) producing only four bands in the gel electropherogram (12.7, 11, 10, and 5 kDa). Liquid chromatography/mass spectrometry analysis reveals a regioselective cleavage in the vicinity of crystallographic CuIIWD/lysozyme interaction sites. Mechanistically, polyoxometalate is critical to position the Cu at the protein surface and limit the generation of oxidative species to the proximity of binding sites. Ultimately, this study outlines the potential of discrete, designable metal oxo clusters as catalysts for the selective modification of proteins through radical mechanisms under non-denaturing conditions.",
publisher = "Springer Nature",
journal = "Nature Communications",
title = "Supplementary material for: Abdelhameed, S. A. M., de Azambuja, F., Vasović, T., Savić, N. D., Ćirković-Veličković, T.,& Parac-Vogt, T. N.. (2023). Regioselective protein oxidative cleavage enabled by enzyme-like recognition of an inorganic metal oxo cluster ligand. in Nature Communications Springer Nature., 14(1), 486. https://doi.org/10.1038/s41467-023-36085-z",
volume = "14",
number = "1",
pages = "486",
url = "https://hdl.handle.net/21.15107/rcub_cherry_5840"
}

Abdelhameed, S. A. M., de Azambuja, F., Vasović, T., Savić, N. D., Ćirković-Veličković, T.,& Parac-Vogt, T. N.. (2023). Supplementary material for: Abdelhameed, S. A. M., de Azambuja, F., Vasović, T., Savić, N. D., Ćirković-Veličković, T.,& Parac-Vogt, T. N.. (2023). Regioselective protein oxidative cleavage enabled by enzyme-like recognition of an inorganic metal oxo cluster ligand. in Nature Communications Springer Nature., 14(1), 486. https://doi.org/10.1038/s41467-023-36085-z. in Nature Communications
Springer Nature., 14(1), 486.
https://hdl.handle.net/21.15107/rcub_cherry_5840

Abdelhameed SAM, de Azambuja F, Vasović T, Savić ND, Ćirković-Veličković T, Parac-Vogt TN. Supplementary material for: Abdelhameed, S. A. M., de Azambuja, F., Vasović, T., Savić, N. D., Ćirković-Veličković, T.,& Parac-Vogt, T. N.. (2023). Regioselective protein oxidative cleavage enabled by enzyme-like recognition of an inorganic metal oxo cluster ligand. in Nature Communications Springer Nature., 14(1), 486. https://doi.org/10.1038/s41467-023-36085-z. in Nature Communications. 2023;14(1):486.
https://hdl.handle.net/21.15107/rcub_cherry_5840 .

Abdelhameed, Shorok A. M., de Azambuja, Francisco, Vasović, Tamara, Savić, Nada D., Ćirković-Veličković, Tanja, Parac-Vogt, Tatjana N., "Supplementary material for: Abdelhameed, S. A. M., de Azambuja, F., Vasović, T., Savić, N. D., Ćirković-Veličković, T.,& Parac-Vogt, T. N.. (2023). Regioselective protein oxidative cleavage enabled by enzyme-like recognition of an inorganic metal oxo cluster ligand. in Nature Communications Springer Nature., 14(1), 486. https://doi.org/10.1038/s41467-023-36085-z" in Nature Communications, 14, no. 1 (2023):486,
https://hdl.handle.net/21.15107/rcub_cherry_5840 .

TY  - CONF
AU  - Krstić-Ristivojević, Maja
AU  - Vasović, Tamara
AU  - Smiljanić, Katarina
AU  - Stanić-Vučinić, Dragana
AU  - Ćirković-Veličković, Tanja
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/5951
AB  - rom over 10 kg in 1960 to over 20 kg in 2014, the yearly per capita consumption of
marine products has increased significantly during the past 50 years. In many nations,
especially those with poor overall protein intake, seafood protein is a crucial component of
the diet1. However, as defined by the European Community shellfish protein tropomyosin
(TPM) is one of the major allergens and major causes of anaphylaxis 2. Although TPM
originating from vertebrates is not considered as an allergen there is evidence that several
fish tropomyosin can be allergenic3. TPM protein is organized of two parallel alpha-helical
molecules which are wound around each other forming a coiled -coil structure2. Although
the degree of similarity between TPM molecules is high, their allergenic potency is
different. Scientists putting a lot of effort into iden tifying and sequencing tropomyosin
isoforms since this information probably explains the intriguing nature of the TPM
molecule. This is a very challenging task given that the differences in mass and pI values
between TPM isoforms are discrete.
TPM was isolated from mussels (Mytilus galloprovincialis), and clams (Venerupis
philippinarum) according to the protocol developed within/and for purposes of the
IMPTOX research project. The obtained “in-house” TPM proteins were resolved using
two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). Isoelectric focusing was
performed on rehydrated Immobiline strips IPG 3-5.6NL 13 cm in Ettan IPGPhor 3 device.
The second dimension was carried out on 12% PAA gels. Upon protein bands excision to
prevent TPM interchain disulfide cross-linking reduction and alkylation of cysteine was
performed. The samples were digested with proteomics-grade trypsin in a 1:50 enzyme-to-
substrate ratio overnight at 37 °C. The obtained peptides were chromatographically
separated using the EASY-nLC II system and analyzed using Orbitrap Exploris 240 mass
spectrometer.
2D-PAGE resolved two isoforms of mussel TPM and up to eight isoforms of clam TPM.
The determined pI value of the dominant isoform of mussel TPM was 4.7 while the
discrete band arising from the second TPM isoform was slightly shifted toward acidic pI
and smaller molecular mass. The determined pI value of the three most dominant clam
TPM isoforms was 4.8, the fourth isoform was slightly shifted toward a more basic pI
value and lower protein molecular weight. The rest of the isoforms were slightly shifted
toward more acidic pI and at a similar molecular weight as the three dominant isoforms.
PB  - Kragujevac : Univerzitet, Prirodno-matematički fakultet
C3  - Zbornik apstrakata, VI Simpozijum Srpskog udruženja za proteomiku (SePA) “Razvoj i primena novih metoda proteomike”, 2. jun 2023. godine, Kragujevac
T1  - Identification of isoforms of shelfish tropomyosin
UR  - https://hdl.handle.net/21.15107/rcub_cherry_5951
ER  - 

@conference{
author = "Krstić-Ristivojević, Maja and Vasović, Tamara and Smiljanić, Katarina and Stanić-Vučinić, Dragana and Ćirković-Veličković, Tanja",
year = "2023",
abstract = "rom over 10 kg in 1960 to over 20 kg in 2014, the yearly per capita consumption of
marine products has increased significantly during the past 50 years. In many nations,
especially those with poor overall protein intake, seafood protein is a crucial component of
the diet1. However, as defined by the European Community shellfish protein tropomyosin
(TPM) is one of the major allergens and major causes of anaphylaxis 2. Although TPM
originating from vertebrates is not considered as an allergen there is evidence that several
fish tropomyosin can be allergenic3. TPM protein is organized of two parallel alpha-helical
molecules which are wound around each other forming a coiled -coil structure2. Although
the degree of similarity between TPM molecules is high, their allergenic potency is
different. Scientists putting a lot of effort into iden tifying and sequencing tropomyosin
isoforms since this information probably explains the intriguing nature of the TPM
molecule. This is a very challenging task given that the differences in mass and pI values
between TPM isoforms are discrete.
TPM was isolated from mussels (Mytilus galloprovincialis), and clams (Venerupis
philippinarum) according to the protocol developed within/and for purposes of the
IMPTOX research project. The obtained “in-house” TPM proteins were resolved using
two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). Isoelectric focusing was
performed on rehydrated Immobiline strips IPG 3-5.6NL 13 cm in Ettan IPGPhor 3 device.
The second dimension was carried out on 12% PAA gels. Upon protein bands excision to
prevent TPM interchain disulfide cross-linking reduction and alkylation of cysteine was
performed. The samples were digested with proteomics-grade trypsin in a 1:50 enzyme-to-
substrate ratio overnight at 37 °C. The obtained peptides were chromatographically
separated using the EASY-nLC II system and analyzed using Orbitrap Exploris 240 mass
spectrometer.
2D-PAGE resolved two isoforms of mussel TPM and up to eight isoforms of clam TPM.
The determined pI value of the dominant isoform of mussel TPM was 4.7 while the
discrete band arising from the second TPM isoform was slightly shifted toward acidic pI
and smaller molecular mass. The determined pI value of the three most dominant clam
TPM isoforms was 4.8, the fourth isoform was slightly shifted toward a more basic pI
value and lower protein molecular weight. The rest of the isoforms were slightly shifted
toward more acidic pI and at a similar molecular weight as the three dominant isoforms.",
publisher = "Kragujevac : Univerzitet, Prirodno-matematički fakultet",
journal = "Zbornik apstrakata, VI Simpozijum Srpskog udruženja za proteomiku (SePA) “Razvoj i primena novih metoda proteomike”, 2. jun 2023. godine, Kragujevac",
title = "Identification of isoforms of shelfish tropomyosin",
url = "https://hdl.handle.net/21.15107/rcub_cherry_5951"
}

Krstić-Ristivojević, M., Vasović, T., Smiljanić, K., Stanić-Vučinić, D.,& Ćirković-Veličković, T.. (2023). Identification of isoforms of shelfish tropomyosin. in Zbornik apstrakata, VI Simpozijum Srpskog udruženja za proteomiku (SePA) “Razvoj i primena novih metoda proteomike”, 2. jun 2023. godine, Kragujevac
Kragujevac : Univerzitet, Prirodno-matematički fakultet..
https://hdl.handle.net/21.15107/rcub_cherry_5951

Krstić-Ristivojević M, Vasović T, Smiljanić K, Stanić-Vučinić D, Ćirković-Veličković T. Identification of isoforms of shelfish tropomyosin. in Zbornik apstrakata, VI Simpozijum Srpskog udruženja za proteomiku (SePA) “Razvoj i primena novih metoda proteomike”, 2. jun 2023. godine, Kragujevac. 2023;.
https://hdl.handle.net/21.15107/rcub_cherry_5951 .

Krstić-Ristivojević, Maja, Vasović, Tamara, Smiljanić, Katarina, Stanić-Vučinić, Dragana, Ćirković-Veličković, Tanja, "Identification of isoforms of shelfish tropomyosin" in Zbornik apstrakata, VI Simpozijum Srpskog udruženja za proteomiku (SePA) “Razvoj i primena novih metoda proteomike”, 2. jun 2023. godine, Kragujevac (2023),
https://hdl.handle.net/21.15107/rcub_cherry_5951 .

TY  - CONF
AU  - Sibinčić, Nikolina
AU  - Krstić-Ristivojević, Maja
AU  - Stojanović, Marijana
AU  - Mladenović Stokanić, Maja
AU  - Vasović, Tamara
AU  - Ćirković-Veličković, Tanja
AU  - Stojadinović, Marija
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6051
AB  - The SARS-CoV-2 nucleocapsid (N) protein plays a significant role in the coronavirus life cycle and participates in a variety of critical events following viral invasion1. In infected patients, high titers of immunoglobulin G (IgG) targeting N protein were detected and correlated with the clinical course of the disease2. Therefore, N protein and anti-N protein IgGs were recognized as important diagnostic indicators of COVID-19 infection in serological and quick antigen tests3. In this study, we optimized the expression of the recombinant form of SARS-CoV-2 N protein in a mammalian cell line HEK293T by comparing the transfection efficiency between Polyethylenimine (PEI) and Calcium Phosphate (CaP) DNA-complexing agents. Transfection potency was tested at different cell confluency and passage number, in several cell culture media, pre-transfection and post-transfection media change and in conditions of reduced serum. Chloroquine and glycerol treatments were included to enhance transfection efficiency as they might inhibit DNA degradation in lysosomes or increase membrane permeability. Protein expression was monitored in cell supernatants up to 7 days post-transfection in dot-bot and Western blot using anti-N protein antibodies. Both transfection methods have shown moderate to relatively high transfection efficiency dependent on the applied conditions, making them affordable and easy to use techniques for recombinant N protein production on a small-scale in adherent mammalian systems. PEI acts as a good delivery system regardless of the presence of the fetal bovine serum (FBS), while CaP transfection is more dependent on the presence of FBS which in turn favors N protein degradation. However, we have optimized both methods to achieve optimal expression of unfragmented N-protein in serum-free conditions. Apart from setting up a cost-effective platform for expression of N protein in mammalian cells, we plan on investigating the mechanisms behind the PEI and CaP non-viral gene delivery systems as there are still some uncertainties in the scientific community.
PB  - Beograde : Serbian Biochemical Society
C3  - "Biochemistry in Biotechnology", Twelfth Conference, International scientific meeting, September 21-23, 2023, Belgrade, Serbia
T1  - Expression of recombinant SARS-CoV-2 nucleocapsid protein in mammalian cells
SP  - 91
EP  - 91
UR  - https://hdl.handle.net/21.15107/rcub_cherry_6051
ER  - 

@conference{
author = "Sibinčić, Nikolina and Krstić-Ristivojević, Maja and Stojanović, Marijana and Mladenović Stokanić, Maja and Vasović, Tamara and Ćirković-Veličković, Tanja and Stojadinović, Marija",
year = "2023",
abstract = "The SARS-CoV-2 nucleocapsid (N) protein plays a significant role in the coronavirus life cycle and participates in a variety of critical events following viral invasion1. In infected patients, high titers of immunoglobulin G (IgG) targeting N protein were detected and correlated with the clinical course of the disease2. Therefore, N protein and anti-N protein IgGs were recognized as important diagnostic indicators of COVID-19 infection in serological and quick antigen tests3. In this study, we optimized the expression of the recombinant form of SARS-CoV-2 N protein in a mammalian cell line HEK293T by comparing the transfection efficiency between Polyethylenimine (PEI) and Calcium Phosphate (CaP) DNA-complexing agents. Transfection potency was tested at different cell confluency and passage number, in several cell culture media, pre-transfection and post-transfection media change and in conditions of reduced serum. Chloroquine and glycerol treatments were included to enhance transfection efficiency as they might inhibit DNA degradation in lysosomes or increase membrane permeability. Protein expression was monitored in cell supernatants up to 7 days post-transfection in dot-bot and Western blot using anti-N protein antibodies. Both transfection methods have shown moderate to relatively high transfection efficiency dependent on the applied conditions, making them affordable and easy to use techniques for recombinant N protein production on a small-scale in adherent mammalian systems. PEI acts as a good delivery system regardless of the presence of the fetal bovine serum (FBS), while CaP transfection is more dependent on the presence of FBS which in turn favors N protein degradation. However, we have optimized both methods to achieve optimal expression of unfragmented N-protein in serum-free conditions. Apart from setting up a cost-effective platform for expression of N protein in mammalian cells, we plan on investigating the mechanisms behind the PEI and CaP non-viral gene delivery systems as there are still some uncertainties in the scientific community.",
publisher = "Beograde : Serbian Biochemical Society",
journal = ""Biochemistry in Biotechnology", Twelfth Conference, International scientific meeting, September 21-23, 2023, Belgrade, Serbia",
title = "Expression of recombinant SARS-CoV-2 nucleocapsid protein in mammalian cells",
pages = "91-91",
url = "https://hdl.handle.net/21.15107/rcub_cherry_6051"
}

Sibinčić, N., Krstić-Ristivojević, M., Stojanović, M., Mladenović Stokanić, M., Vasović, T., Ćirković-Veličković, T.,& Stojadinović, M.. (2023). Expression of recombinant SARS-CoV-2 nucleocapsid protein in mammalian cells. in "Biochemistry in Biotechnology", Twelfth Conference, International scientific meeting, September 21-23, 2023, Belgrade, Serbia
Beograde : Serbian Biochemical Society., 91-91.
https://hdl.handle.net/21.15107/rcub_cherry_6051

Sibinčić N, Krstić-Ristivojević M, Stojanović M, Mladenović Stokanić M, Vasović T, Ćirković-Veličković T, Stojadinović M. Expression of recombinant SARS-CoV-2 nucleocapsid protein in mammalian cells. in "Biochemistry in Biotechnology", Twelfth Conference, International scientific meeting, September 21-23, 2023, Belgrade, Serbia. 2023;:91-91.
https://hdl.handle.net/21.15107/rcub_cherry_6051 .

Sibinčić, Nikolina, Krstić-Ristivojević, Maja, Stojanović, Marijana, Mladenović Stokanić, Maja, Vasović, Tamara, Ćirković-Veličković, Tanja, Stojadinović, Marija, "Expression of recombinant SARS-CoV-2 nucleocapsid protein in mammalian cells" in "Biochemistry in Biotechnology", Twelfth Conference, International scientific meeting, September 21-23, 2023, Belgrade, Serbia (2023):91-91,
https://hdl.handle.net/21.15107/rcub_cherry_6051 .

TY  - GEN
AU  - Ćirković-Veličković, Tanja
AU  - Gnjatović, Marija
AU  - Ćujić, Danica
AU  - Todorović, Aleksandra
AU  - Stanić-Vučinić, Dragana
AU  - Đukić, Teodora
AU  - Mladenović, Maja
AU  - Vasović, Tamara
AU  - Stojadinović, Marija
AU  - Krstić-Ristivojević, Maja
AU  - Jovanović, Vesna
AU  - Simović, Ana
AU  - Radosavljević, Jelena
AU  - Aćimović, Jelena M.
AU  - Radomirović, Mirjana Ž.
AU  - Stojanović, Marijana
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6014
AB  - Нови корона вирус (SARS CoV-2) који се појавио у Вухану 2019. године припада групи једноланчаних РНК вируса [1]. Представља нови инфективни агенс за хуману популацију и веома је брзо детектован у великом броју земаља. Узрочник је респираторних инфекција које могу да буду праћене и веома тешком клиничком сликом. Брзо ширење, одсуство имунитета на овај вирус и одсуство поузданих тестова за детекцију вируса у тренутку избијања пандемије су болест изазвану овим вирусом брзо претворили у здравствени и друштвени проблем највишег приоритета на глобалном нивоу. Иако су највеће биотехнолошке компаније убрзано почеле са развојем и масовном производњом дијагностичких тестова и вакцина, њихова доступност у тренуцима највеће потражње је и даље недовољна, а цене истих су лимитирајући фактор за бољу контролу болести и ширења пандемије [2]. Развој сопствених и одржива производња тестова и вакцина за COVID-19 су од великог друштвеног значаја. Важан предуслов за одрживу производњу тестова је доступност рекомбинантних антигена вируса и могућност производње истих на великој скали за потребе производње домаћих тестова. Овим техничким решењем се описује добијање два кључна антигена новог корона вируса рекомбинантном технологијом и њихова примена у серолошком ЕЛИСА тесту који производи Институт за примену нуклеарне енергије, ИНЕП, као и за добијање реагенаса за детекцију антигена новог корона вируса (специфичних антитела). У првој фази, оптимизоване су секвенце протеина које су подигле осетљивост постојећих серолошких тестова. Иновативност нашег приступа се огледа и у разрађеним експерименталним протоколима за добијање рекомбинантних протеина нуклеокапсида на великој скали, као и у солубилној форми, што олакшава поступак пречишћавања. Избор фрагмента нуклеокапсида који се хетеролого експримира у солубилној форми, а специфично детектује антитела и генерише јак имуни одговор током имунизације животиња (имуногеност) на основу прегледа познатих епитопских секвенци је кључна иновација овог техничког решења. Ово је први пример успешно примењеног рекомбинатног протеина произведеног у Србији у дијагностичком тесту који је регистрован
код Агенције за лекове и медицинска средства Републике Србије (број решења 515-02-02370-21-002), а који је примену нашао и на међународном нивоу.
T1  - Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus
UR  - https://hdl.handle.net/21.15107/rcub_cherry_6014
ER  - 

@misc{
author = "Ćirković-Veličković, Tanja and Gnjatović, Marija and Ćujić, Danica and Todorović, Aleksandra and Stanić-Vučinić, Dragana and Đukić, Teodora and Mladenović, Maja and Vasović, Tamara and Stojadinović, Marija and Krstić-Ristivojević, Maja and Jovanović, Vesna and Simović, Ana and Radosavljević, Jelena and Aćimović, Jelena M. and Radomirović, Mirjana Ž. and Stojanović, Marijana",
year = "2023",
abstract = "Нови корона вирус (SARS CoV-2) који се појавио у Вухану 2019. године припада групи једноланчаних РНК вируса [1]. Представља нови инфективни агенс за хуману популацију и веома је брзо детектован у великом броју земаља. Узрочник је респираторних инфекција које могу да буду праћене и веома тешком клиничком сликом. Брзо ширење, одсуство имунитета на овај вирус и одсуство поузданих тестова за детекцију вируса у тренутку избијања пандемије су болест изазвану овим вирусом брзо претворили у здравствени и друштвени проблем највишег приоритета на глобалном нивоу. Иако су највеће биотехнолошке компаније убрзано почеле са развојем и масовном производњом дијагностичких тестова и вакцина, њихова доступност у тренуцима највеће потражње је и даље недовољна, а цене истих су лимитирајући фактор за бољу контролу болести и ширења пандемије [2]. Развој сопствених и одржива производња тестова и вакцина за COVID-19 су од великог друштвеног значаја. Важан предуслов за одрживу производњу тестова је доступност рекомбинантних антигена вируса и могућност производње истих на великој скали за потребе производње домаћих тестова. Овим техничким решењем се описује добијање два кључна антигена новог корона вируса рекомбинантном технологијом и њихова примена у серолошком ЕЛИСА тесту који производи Институт за примену нуклеарне енергије, ИНЕП, као и за добијање реагенаса за детекцију антигена новог корона вируса (специфичних антитела). У првој фази, оптимизоване су секвенце протеина које су подигле осетљивост постојећих серолошких тестова. Иновативност нашег приступа се огледа и у разрађеним експерименталним протоколима за добијање рекомбинантних протеина нуклеокапсида на великој скали, као и у солубилној форми, што олакшава поступак пречишћавања. Избор фрагмента нуклеокапсида који се хетеролого експримира у солубилној форми, а специфично детектује антитела и генерише јак имуни одговор током имунизације животиња (имуногеност) на основу прегледа познатих епитопских секвенци је кључна иновација овог техничког решења. Ово је први пример успешно примењеног рекомбинатног протеина произведеног у Србији у дијагностичком тесту који је регистрован
код Агенције за лекове и медицинска средства Републике Србије (број решења 515-02-02370-21-002), а који је примену нашао и на међународном нивоу.",
title = "Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus",
url = "https://hdl.handle.net/21.15107/rcub_cherry_6014"
}

Ćirković-Veličković, T., Gnjatović, M., Ćujić, D., Todorović, A., Stanić-Vučinić, D., Đukić, T., Mladenović, M., Vasović, T., Stojadinović, M., Krstić-Ristivojević, M., Jovanović, V., Simović, A., Radosavljević, J., Aćimović, J. M., Radomirović, M. Ž.,& Stojanović, M.. (2023). Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus. .
https://hdl.handle.net/21.15107/rcub_cherry_6014

Ćirković-Veličković T, Gnjatović M, Ćujić D, Todorović A, Stanić-Vučinić D, Đukić T, Mladenović M, Vasović T, Stojadinović M, Krstić-Ristivojević M, Jovanović V, Simović A, Radosavljević J, Aćimović JM, Radomirović MŽ, Stojanović M. Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus. 2023;.
https://hdl.handle.net/21.15107/rcub_cherry_6014 .

Ćirković-Veličković, Tanja, Gnjatović, Marija, Ćujić, Danica, Todorović, Aleksandra, Stanić-Vučinić, Dragana, Đukić, Teodora, Mladenović, Maja, Vasović, Tamara, Stojadinović, Marija, Krstić-Ristivojević, Maja, Jovanović, Vesna, Simović, Ana, Radosavljević, Jelena, Aćimović, Jelena M., Radomirović, Mirjana Ž., Stojanović, Marijana, "Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus" (2023),
https://hdl.handle.net/21.15107/rcub_cherry_6014 .

TY  - CONF
AU  - Radomirović, Mirjana Ž.
AU  - Bićanin, Maša
AU  - Udovički, Božidar
AU  - Krstić-Ristivojević, Maja
AU  - Đukić, Teodora
AU  - Vasović, Tamara
AU  - Jovanović, Vesna
AU  - Stanić-Vučinić, Dragana
AU  - Rajković, Andreja
AU  - Ćirković-Veličković, Tanja
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6021
AB  - Accurately diagnosing people with suspected SARS-CoV-2 infection is essential to help manage COVID-19. Currently available SARS-CoV-2 diagnostics detect either RNA of the virus by RT-PCR or the presence of viral antigens in biological fluids by ELISA or similar techniques. Low sensitivity of antigen tests could lead to the risk of false negative results. Therefore, this study aimed to develop a highly sensitive immuno-PCR method for quantifying SARS-CoV-2 nucleocapsid (N) protein that combines the specificity of sandwich ELISA with the sensitivity of PCR. Recombinant N protein fragment was produced in E. coli as an expression system and purified using immobilized metal ion affinity chromatography. The antibodies against the N protein were raised in rabbits and mice. High-affinity polyclonal mice and rabbit N protein-specific antisera were purified using ammonium sulfate precipitation and used to develop sandwich ELISA for the quantification of N protein. Mice polyclonal serum was used as a capture for N protein. N
protein bound to mice antibodies was detected with rabbit polyclonal sera. A double-stranded amino-DNA molecule of 77 base pairs was PCR-synthesized, covalently conjugated to a secondary goat anti-rabbit antibody and subsequently amplified and quantified by real-time PCR. The results were compared to analogous sandwich ELISA consisting of alkaline phosphatase-labeled goat anti-rabbit antibody. The sensitivity of immuno-PCR for quantification of N protein was increased by up to 7-fold compared to analogous ELISA, having a limit of detection of 92 pg/mL and a limit of quantification of 840 pg/mL. The developed immuno-PCR method thus has the potential to be used as a new antigen test for COVID-19 and beyond.
PB  - Federation of European Biochemical Societies, Wiley
C3  - The 47th FEBS Congress, 8th-12th July, 2023. In: FEBS Open Bio, 13: Suppl. 2
T1  - Development of immuno-PCR for sensitive quantification of SARS-CoV-2 nucleocapsid protein
SP  - 44
EP  - 44
UR  - https://hdl.handle.net/21.15107/rcub_cherry_6021
ER  - 

@conference{
author = "Radomirović, Mirjana Ž. and Bićanin, Maša and Udovički, Božidar and Krstić-Ristivojević, Maja and Đukić, Teodora and Vasović, Tamara and Jovanović, Vesna and Stanić-Vučinić, Dragana and Rajković, Andreja and Ćirković-Veličković, Tanja",
year = "2023",
abstract = "Accurately diagnosing people with suspected SARS-CoV-2 infection is essential to help manage COVID-19. Currently available SARS-CoV-2 diagnostics detect either RNA of the virus by RT-PCR or the presence of viral antigens in biological fluids by ELISA or similar techniques. Low sensitivity of antigen tests could lead to the risk of false negative results. Therefore, this study aimed to develop a highly sensitive immuno-PCR method for quantifying SARS-CoV-2 nucleocapsid (N) protein that combines the specificity of sandwich ELISA with the sensitivity of PCR. Recombinant N protein fragment was produced in E. coli as an expression system and purified using immobilized metal ion affinity chromatography. The antibodies against the N protein were raised in rabbits and mice. High-affinity polyclonal mice and rabbit N protein-specific antisera were purified using ammonium sulfate precipitation and used to develop sandwich ELISA for the quantification of N protein. Mice polyclonal serum was used as a capture for N protein. N
protein bound to mice antibodies was detected with rabbit polyclonal sera. A double-stranded amino-DNA molecule of 77 base pairs was PCR-synthesized, covalently conjugated to a secondary goat anti-rabbit antibody and subsequently amplified and quantified by real-time PCR. The results were compared to analogous sandwich ELISA consisting of alkaline phosphatase-labeled goat anti-rabbit antibody. The sensitivity of immuno-PCR for quantification of N protein was increased by up to 7-fold compared to analogous ELISA, having a limit of detection of 92 pg/mL and a limit of quantification of 840 pg/mL. The developed immuno-PCR method thus has the potential to be used as a new antigen test for COVID-19 and beyond.",
publisher = "Federation of European Biochemical Societies, Wiley",
journal = "The 47th FEBS Congress, 8th-12th July, 2023. In: FEBS Open Bio, 13: Suppl. 2",
title = "Development of immuno-PCR for sensitive quantification of SARS-CoV-2 nucleocapsid protein",
pages = "44-44",
url = "https://hdl.handle.net/21.15107/rcub_cherry_6021"
}

Radomirović, M. Ž., Bićanin, M., Udovički, B., Krstić-Ristivojević, M., Đukić, T., Vasović, T., Jovanović, V., Stanić-Vučinić, D., Rajković, A.,& Ćirković-Veličković, T.. (2023). Development of immuno-PCR for sensitive quantification of SARS-CoV-2 nucleocapsid protein. in The 47th FEBS Congress, 8th-12th July, 2023. In: FEBS Open Bio, 13: Suppl. 2
Federation of European Biochemical Societies, Wiley., 44-44.
https://hdl.handle.net/21.15107/rcub_cherry_6021

Radomirović MŽ, Bićanin M, Udovički B, Krstić-Ristivojević M, Đukić T, Vasović T, Jovanović V, Stanić-Vučinić D, Rajković A, Ćirković-Veličković T. Development of immuno-PCR for sensitive quantification of SARS-CoV-2 nucleocapsid protein. in The 47th FEBS Congress, 8th-12th July, 2023. In: FEBS Open Bio, 13: Suppl. 2. 2023;:44-44.
https://hdl.handle.net/21.15107/rcub_cherry_6021 .

Radomirović, Mirjana Ž., Bićanin, Maša, Udovički, Božidar, Krstić-Ristivojević, Maja, Đukić, Teodora, Vasović, Tamara, Jovanović, Vesna, Stanić-Vučinić, Dragana, Rajković, Andreja, Ćirković-Veličković, Tanja, "Development of immuno-PCR for sensitive quantification of SARS-CoV-2 nucleocapsid protein" in The 47th FEBS Congress, 8th-12th July, 2023. In: FEBS Open Bio, 13: Suppl. 2 (2023):44-44,
https://hdl.handle.net/21.15107/rcub_cherry_6021 .

TY  - CONF
AU  - Gligorijević, Nikola
AU  - Stanić-Vučinić, Dragana
AU  - Mutić, Tamara
AU  - Lujić, Tamara
AU  - Ćirković-Veličković, Tanja
PY  - 2023
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/5909
AB  - Microplastic represents one of the major types of pollutants in modern era. Over several years of research in the field of microplastic,
there are still many unknown gaps, including the effects and mechanisms of action of these particles on human
health. Studies in this field conducted experiments on cells and human tissues or animals like rats and mice. While these
studies suggest the toxic effects of microplastic, it is not clear if concentrations used for exposure are relevant for humans.
Also, most of the studies used spherical polystyrene, which does not reflect well the diversity of microplastic particles found
in nature. Another gap is lack of studies describing direct interactions of microplastics and proteins. While it is generally
known that proteins form corona around microplastic particles, affinity studies and consequences on protein structure are
usually missing.
The aim of this work was to analyze interaction of a major egg white protein and allergen, ovalbumin to several to microplastic
particles, including polystyrene (PS) of 120 and 500 μm in size and polyethylene terephthalate (PET) of 120 μm in
size. Binding affinity was determined at both acidic, pH 3 and neutral, pH 7 conditions, at the room temperature, by measuring
bulk ovalbumin concentration in supernatants at the equilibrium time. Several binding models, including Langmuir,
Freundlich, Redlich–Peterson and Guggenheim-Anderson-de Boer (GAB), were used to determine binding parameters.
The formation of soft and hard corona was analyzed according to the published protocol [1]. Structural analysis was performed
using near and far-UV CD spectrometry.
Obtained results showed that ovalbumin binds to both PS and PET. All binding models indicated that ovalbumin binds
with higher affinity to tested microplastics on pH 3, compared to pH 7, with the highest affinity being calculated for PS 120
μm. Further analysis showed that ovalbumin forms both soft and hard corona onto the surface of all three microplastics.
Structural alterations of ovalbumin as a consequence of its interaction with microplastic was shown to be both pH and
microplastic type dependent. Also, more pronounced effect on its tertiary structure was observed, compared to secondary.
At pH3, tertiary structure of bulk ovalbumin becomes destabilized, especially in the presence of PET 120 μm and PS 500
μm, while at pH 7, structural stabilization is observed, especially in the presence of PS 120 μm.
Considering that the microplastic was discovered in eggs [2], obtained results suggest that direct interactions of native
ovalbumin with microplastic particles could have influence on its structure and thus affect its techno-functional properties.
Acknowledgments: This project has received funding from the European Union’s Horizon 2020 research and innovation programme
under grant agreement No 96517.
References:
[1] D. Magrì, P. Sánchez-Moreno, G. Caputo, F. Gatto, M. Veronesi, G. Bardi, T. Catelani, D. Guarnieri, A. Athanassiou, P.P. Pompa, D.
Fragouli, ACS Nano, 12 (2018) 7690-7700.
[2] Q. Liu, Z. Chen, Y. Chen, F. Yang, W. Yao, Y. Xie, Food Chemistry, 397 (2022) 133771
PB  - Belgrade : Serbian Chemical Society
C3  - Book of Abstracts of the XXII EuroFoodChem Congress, Belgrade, Serbia, 14-16 June 2023
T1  - Binding and corona formation of ovalbumin to polystyrene and polyethylene terephthalate microplastics under neutral and acidic conditions
SP  - 137
EP  - 137
UR  - https://hdl.handle.net/21.15107/rcub_cherry_5909
ER  - 

@conference{
author = "Gligorijević, Nikola and Stanić-Vučinić, Dragana and Mutić, Tamara and Lujić, Tamara and Ćirković-Veličković, Tanja",
year = "2023",
abstract = "Microplastic represents one of the major types of pollutants in modern era. Over several years of research in the field of microplastic,
there are still many unknown gaps, including the effects and mechanisms of action of these particles on human
health. Studies in this field conducted experiments on cells and human tissues or animals like rats and mice. While these
studies suggest the toxic effects of microplastic, it is not clear if concentrations used for exposure are relevant for humans.
Also, most of the studies used spherical polystyrene, which does not reflect well the diversity of microplastic particles found
in nature. Another gap is lack of studies describing direct interactions of microplastics and proteins. While it is generally
known that proteins form corona around microplastic particles, affinity studies and consequences on protein structure are
usually missing.
The aim of this work was to analyze interaction of a major egg white protein and allergen, ovalbumin to several to microplastic
particles, including polystyrene (PS) of 120 and 500 μm in size and polyethylene terephthalate (PET) of 120 μm in
size. Binding affinity was determined at both acidic, pH 3 and neutral, pH 7 conditions, at the room temperature, by measuring
bulk ovalbumin concentration in supernatants at the equilibrium time. Several binding models, including Langmuir,
Freundlich, Redlich–Peterson and Guggenheim-Anderson-de Boer (GAB), were used to determine binding parameters.
The formation of soft and hard corona was analyzed according to the published protocol [1]. Structural analysis was performed
using near and far-UV CD spectrometry.
Obtained results showed that ovalbumin binds to both PS and PET. All binding models indicated that ovalbumin binds
with higher affinity to tested microplastics on pH 3, compared to pH 7, with the highest affinity being calculated for PS 120
μm. Further analysis showed that ovalbumin forms both soft and hard corona onto the surface of all three microplastics.
Structural alterations of ovalbumin as a consequence of its interaction with microplastic was shown to be both pH and
microplastic type dependent. Also, more pronounced effect on its tertiary structure was observed, compared to secondary.
At pH3, tertiary structure of bulk ovalbumin becomes destabilized, especially in the presence of PET 120 μm and PS 500
μm, while at pH 7, structural stabilization is observed, especially in the presence of PS 120 μm.
Considering that the microplastic was discovered in eggs [2], obtained results suggest that direct interactions of native
ovalbumin with microplastic particles could have influence on its structure and thus affect its techno-functional properties.
Acknowledgments: This project has received funding from the European Union’s Horizon 2020 research and innovation programme
under grant agreement No 96517.
References:
[1] D. Magrì, P. Sánchez-Moreno, G. Caputo, F. Gatto, M. Veronesi, G. Bardi, T. Catelani, D. Guarnieri, A. Athanassiou, P.P. Pompa, D.
Fragouli, ACS Nano, 12 (2018) 7690-7700.
[2] Q. Liu, Z. Chen, Y. Chen, F. Yang, W. Yao, Y. Xie, Food Chemistry, 397 (2022) 133771",
publisher = "Belgrade : Serbian Chemical Society",
journal = "Book of Abstracts of the XXII EuroFoodChem Congress, Belgrade, Serbia, 14-16 June 2023",
title = "Binding and corona formation of ovalbumin to polystyrene and polyethylene terephthalate microplastics under neutral and acidic conditions",
pages = "137-137",
url = "https://hdl.handle.net/21.15107/rcub_cherry_5909"
}

Gligorijević, N., Stanić-Vučinić, D., Mutić, T., Lujić, T.,& Ćirković-Veličković, T.. (2023). Binding and corona formation of ovalbumin to polystyrene and polyethylene terephthalate microplastics under neutral and acidic conditions. in Book of Abstracts of the XXII EuroFoodChem Congress, Belgrade, Serbia, 14-16 June 2023
Belgrade : Serbian Chemical Society., 137-137.
https://hdl.handle.net/21.15107/rcub_cherry_5909

Gligorijević N, Stanić-Vučinić D, Mutić T, Lujić T, Ćirković-Veličković T. Binding and corona formation of ovalbumin to polystyrene and polyethylene terephthalate microplastics under neutral and acidic conditions. in Book of Abstracts of the XXII EuroFoodChem Congress, Belgrade, Serbia, 14-16 June 2023. 2023;:137-137.
https://hdl.handle.net/21.15107/rcub_cherry_5909 .

Gligorijević, Nikola, Stanić-Vučinić, Dragana, Mutić, Tamara, Lujić, Tamara, Ćirković-Veličković, Tanja, "Binding and corona formation of ovalbumin to polystyrene and polyethylene terephthalate microplastics under neutral and acidic conditions" in Book of Abstracts of the XXII EuroFoodChem Congress, Belgrade, Serbia, 14-16 June 2023 (2023):137-137,
https://hdl.handle.net/21.15107/rcub_cherry_5909 .