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AB  - Research data for the unpublished paper: Ovalbumin interaction with polystyrene and polyethylene terephthalate microplastics alters its structural properties.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abstract = "Research data for the unpublished paper: Ovalbumin interaction with polystyrene and polyethylene terephthalate microplastics alters its structural properties.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  - 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  - 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  - 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  - 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  - JOUR
AU  - Vujatović, Tamara B.
AU  - Vitorović-Todorović, Maja D.
AU  - Cvijetić, Ilija
AU  - Vasović, Tamara
AU  - Nikolić, Milan
AU  - Novaković, Irena T.
AU  - Bjelogrlić, Snežana K.
PY  - 2022
UR  - https://www.sciencedirect.com/science/article/pii/S0022286021018287
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/4761
AB  - In the present work, the α,β-double bond of the aroylacrylic acid phenylamides was suitably modified to optimize the toxicity–antiproliferative activity ratio of the resulting compounds 1–5. The phenylamides were modified by Michael's addition of suitably chosen piperidine-containing fragments: 1-amino-N-benzylpiperidine (a1), 4-benzylpiperidine (a2), and N,N-dimethyl-N-[2-(1-piperazinyl)-ethyl]amine (a3). The compounds exerted micromolar activity toward three cancer cell lines, A549, LoVo, and Skov-3, causing apoptotic cell death. It was shown that the nature of the cyclic amine moiety at position C2 of the compounds is probably the primary determinant of anticancer activity toward tested cell lines and the acute toxicity toward brine shrimp (Artemia salina). The majority of compounds revealed the ability to vigorously induce mitochondrial superoxide anion generation in all treated cell lines, which together with cell cycle arrest at the S phase and activation of intrinsic caspase cascade, indicates the possibility that apoptosis was triggered due to irreparable chromosomal damage by acute oxidative stress. Two derivatives also exerted significant antibacterial activity with one order of magnitude higher potency than chloramphenicol in most of the investigated bacterial strains. Also, the drug-like properties for all compounds were estimated by available software tools.
PB  - Elsevier
T2  - Journal of Molecular Structure
T1  - Novel derivatives of aroylacrylic acid phenylamides as inducers of apoptosis through the ROS-mediated pathway in several cancer cell lines
VL  - 1250
SP  - 131702
DO  - 10.1016/j.molstruc.2021.131702
ER  - 

@article{
author = "Vujatović, Tamara B. and Vitorović-Todorović, Maja D. and Cvijetić, Ilija and Vasović, Tamara and Nikolić, Milan and Novaković, Irena T. and Bjelogrlić, Snežana K.",
year = "2022",
abstract = "In the present work, the α,β-double bond of the aroylacrylic acid phenylamides was suitably modified to optimize the toxicity–antiproliferative activity ratio of the resulting compounds 1–5. The phenylamides were modified by Michael's addition of suitably chosen piperidine-containing fragments: 1-amino-N-benzylpiperidine (a1), 4-benzylpiperidine (a2), and N,N-dimethyl-N-[2-(1-piperazinyl)-ethyl]amine (a3). The compounds exerted micromolar activity toward three cancer cell lines, A549, LoVo, and Skov-3, causing apoptotic cell death. It was shown that the nature of the cyclic amine moiety at position C2 of the compounds is probably the primary determinant of anticancer activity toward tested cell lines and the acute toxicity toward brine shrimp (Artemia salina). The majority of compounds revealed the ability to vigorously induce mitochondrial superoxide anion generation in all treated cell lines, which together with cell cycle arrest at the S phase and activation of intrinsic caspase cascade, indicates the possibility that apoptosis was triggered due to irreparable chromosomal damage by acute oxidative stress. Two derivatives also exerted significant antibacterial activity with one order of magnitude higher potency than chloramphenicol in most of the investigated bacterial strains. Also, the drug-like properties for all compounds were estimated by available software tools.",
publisher = "Elsevier",
journal = "Journal of Molecular Structure",
title = "Novel derivatives of aroylacrylic acid phenylamides as inducers of apoptosis through the ROS-mediated pathway in several cancer cell lines",
volume = "1250",
pages = "131702",
doi = "10.1016/j.molstruc.2021.131702"
}

Vujatović, T. B., Vitorović-Todorović, M. D., Cvijetić, I., Vasović, T., Nikolić, M., Novaković, I. T.,& Bjelogrlić, S. K.. (2022). Novel derivatives of aroylacrylic acid phenylamides as inducers of apoptosis through the ROS-mediated pathway in several cancer cell lines. in Journal of Molecular Structure
Elsevier., 1250, 131702.
https://doi.org/10.1016/j.molstruc.2021.131702

Vujatović TB, Vitorović-Todorović MD, Cvijetić I, Vasović T, Nikolić M, Novaković IT, Bjelogrlić SK. Novel derivatives of aroylacrylic acid phenylamides as inducers of apoptosis through the ROS-mediated pathway in several cancer cell lines. in Journal of Molecular Structure. 2022;1250:131702.
doi:10.1016/j.molstruc.2021.131702 .

Vujatović, Tamara B., Vitorović-Todorović, Maja D., Cvijetić, Ilija, Vasović, Tamara, Nikolić, Milan, Novaković, Irena T., Bjelogrlić, Snežana K., "Novel derivatives of aroylacrylic acid phenylamides as inducers of apoptosis through the ROS-mediated pathway in several cancer cell lines" in Journal of Molecular Structure, 1250 (2022):131702,
https://doi.org/10.1016/j.molstruc.2021.131702 . .

TY  - DATA
AU  - Vujatović, Tamara B.
AU  - Vitorović-Todorović, Maja D.
AU  - Cvijetić, Ilija
AU  - Vasović, Tamara
AU  - Nikolić, Milan
AU  - Novaković, Irena T.
AU  - Bjelogrlić, Snežana K.
PY  - 2022
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/4762
PB  - Elsevier
T2  - Journal of Molecular Structure
T1  - Supplementary data for the article: Vujatović, T. B.; Vitorović-Todorović, M. D.; Cvijetić, I.; Vasović, T.; Nikolić, M. R.; Novaković, I.; Bjelogrlić, S. Novel Derivatives of Aroylacrylic Acid Phenylamides as Inducers of Apoptosis through the ROS-Mediated Pathway in Several Cancer Cell Lines. Journal of Molecular Structure 2022, 1250, 131702. https://doi.org/10.1016/j.molstruc.2021.131702.
UR  - https://hdl.handle.net/21.15107/rcub_cherry_4762
ER  - 

@misc{
author = "Vujatović, Tamara B. and Vitorović-Todorović, Maja D. and Cvijetić, Ilija and Vasović, Tamara and Nikolić, Milan and Novaković, Irena T. and Bjelogrlić, Snežana K.",
year = "2022",
publisher = "Elsevier",
journal = "Journal of Molecular Structure",
title = "Supplementary data for the article: Vujatović, T. B.; Vitorović-Todorović, M. D.; Cvijetić, I.; Vasović, T.; Nikolić, M. R.; Novaković, I.; Bjelogrlić, S. Novel Derivatives of Aroylacrylic Acid Phenylamides as Inducers of Apoptosis through the ROS-Mediated Pathway in Several Cancer Cell Lines. Journal of Molecular Structure 2022, 1250, 131702. https://doi.org/10.1016/j.molstruc.2021.131702.",
url = "https://hdl.handle.net/21.15107/rcub_cherry_4762"
}

Vujatović, T. B., Vitorović-Todorović, M. D., Cvijetić, I., Vasović, T., Nikolić, M., Novaković, I. T.,& Bjelogrlić, S. K.. (2022). Supplementary data for the article: Vujatović, T. B.; Vitorović-Todorović, M. D.; Cvijetić, I.; Vasović, T.; Nikolić, M. R.; Novaković, I.; Bjelogrlić, S. Novel Derivatives of Aroylacrylic Acid Phenylamides as Inducers of Apoptosis through the ROS-Mediated Pathway in Several Cancer Cell Lines. Journal of Molecular Structure 2022, 1250, 131702. https://doi.org/10.1016/j.molstruc.2021.131702.. in Journal of Molecular Structure
Elsevier..
https://hdl.handle.net/21.15107/rcub_cherry_4762

Vujatović TB, Vitorović-Todorović MD, Cvijetić I, Vasović T, Nikolić M, Novaković IT, Bjelogrlić SK. Supplementary data for the article: Vujatović, T. B.; Vitorović-Todorović, M. D.; Cvijetić, I.; Vasović, T.; Nikolić, M. R.; Novaković, I.; Bjelogrlić, S. Novel Derivatives of Aroylacrylic Acid Phenylamides as Inducers of Apoptosis through the ROS-Mediated Pathway in Several Cancer Cell Lines. Journal of Molecular Structure 2022, 1250, 131702. https://doi.org/10.1016/j.molstruc.2021.131702.. in Journal of Molecular Structure. 2022;.
https://hdl.handle.net/21.15107/rcub_cherry_4762 .

Vujatović, Tamara B., Vitorović-Todorović, Maja D., Cvijetić, Ilija, Vasović, Tamara, Nikolić, Milan, Novaković, Irena T., Bjelogrlić, Snežana K., "Supplementary data for the article: Vujatović, T. B.; Vitorović-Todorović, M. D.; Cvijetić, I.; Vasović, T.; Nikolić, M. R.; Novaković, I.; Bjelogrlić, S. Novel Derivatives of Aroylacrylic Acid Phenylamides as Inducers of Apoptosis through the ROS-Mediated Pathway in Several Cancer Cell Lines. Journal of Molecular Structure 2022, 1250, 131702. https://doi.org/10.1016/j.molstruc.2021.131702." in Journal of Molecular Structure (2022),
https://hdl.handle.net/21.15107/rcub_cherry_4762 .

TY  - JOUR
AU  - Đukić, Teodora
AU  - Mladenović, Maja
AU  - Stanić-Vučinić, Dragana
AU  - Radosavljević, Jelena
AU  - Smiljanić, Katarina
AU  - Sabljić, Ljiljana
AU  - Dević, Marija
AU  - Ćujić, Danica R.
AU  - Vasović, Tamara
AU  - Simović, Ana
AU  - Radomirović, Mirjana Ž.
AU  - Ćirković-Veličković, Tanja
PY  - 2021
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/4330
AB  - Serological testing is important method for diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Nucleocapsid (N) protein is the most abundant virus derived protein and strong immunogen. We aimed to find its efficient, low-cost production.

SARS-CoV-2 recombinant fragment of nucleocapsid protein (rfNP; 58–419 aa) was expressed in E. coli in soluble form, purified and characterized biochemically and immunologically.

Purified rfNP has secondary structure of full-length recombinant N protein, with high percentage of disordered structure (34.2%) and of β-sheet (40.7%). rfNP was tested in immunoblot using sera of COVID-19 convalescent patients. ELISA was optimized with sera of RT-PCR confirmed positive symptomatic patients and healthy individuals. IgG detection sensitivity was 96% (47/50) and specificity 97% (67/68), while IgM detection was slightly lower (94% and 96.5%, respectively).

Cost-effective approach for soluble recombinant N protein fragment production was developed, with reliable IgG and IgM antibodies detection of SARS-CoV-2 infection.
PB  - Elsevier
T2  - Virology journal
T1  - Expression, purification and immunological characterization of recombinant nucleocapsid protein fragment from SARS-CoV-2
VL  - 557
SP  - 15
EP  - 22
DO  - 10.1016/j.virol.2021.01.004
ER  - 

@article{
author = "Đukić, Teodora and Mladenović, Maja and Stanić-Vučinić, Dragana and Radosavljević, Jelena and Smiljanić, Katarina and Sabljić, Ljiljana and Dević, Marija and Ćujić, Danica R. and Vasović, Tamara and Simović, Ana and Radomirović, Mirjana Ž. and Ćirković-Veličković, Tanja",
year = "2021",
abstract = "Serological testing is important method for diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Nucleocapsid (N) protein is the most abundant virus derived protein and strong immunogen. We aimed to find its efficient, low-cost production.

SARS-CoV-2 recombinant fragment of nucleocapsid protein (rfNP; 58–419 aa) was expressed in E. coli in soluble form, purified and characterized biochemically and immunologically.

Purified rfNP has secondary structure of full-length recombinant N protein, with high percentage of disordered structure (34.2%) and of β-sheet (40.7%). rfNP was tested in immunoblot using sera of COVID-19 convalescent patients. ELISA was optimized with sera of RT-PCR confirmed positive symptomatic patients and healthy individuals. IgG detection sensitivity was 96% (47/50) and specificity 97% (67/68), while IgM detection was slightly lower (94% and 96.5%, respectively).

Cost-effective approach for soluble recombinant N protein fragment production was developed, with reliable IgG and IgM antibodies detection of SARS-CoV-2 infection.",
publisher = "Elsevier",
journal = "Virology journal",
title = "Expression, purification and immunological characterization of recombinant nucleocapsid protein fragment from SARS-CoV-2",
volume = "557",
pages = "15-22",
doi = "10.1016/j.virol.2021.01.004"
}

Đukić, T., Mladenović, M., Stanić-Vučinić, D., Radosavljević, J., Smiljanić, K., Sabljić, L., Dević, M., Ćujić, D. R., Vasović, T., Simović, A., Radomirović, M. Ž.,& Ćirković-Veličković, T.. (2021). Expression, purification and immunological characterization of recombinant nucleocapsid protein fragment from SARS-CoV-2. in Virology journal
Elsevier., 557, 15-22.
https://doi.org/10.1016/j.virol.2021.01.004

Đukić T, Mladenović M, Stanić-Vučinić D, Radosavljević J, Smiljanić K, Sabljić L, Dević M, Ćujić DR, Vasović T, Simović A, Radomirović MŽ, Ćirković-Veličković T. Expression, purification and immunological characterization of recombinant nucleocapsid protein fragment from SARS-CoV-2. in Virology journal. 2021;557:15-22.
doi:10.1016/j.virol.2021.01.004 .

Đukić, Teodora, Mladenović, Maja, Stanić-Vučinić, Dragana, Radosavljević, Jelena, Smiljanić, Katarina, Sabljić, Ljiljana, Dević, Marija, Ćujić, Danica R., Vasović, Tamara, Simović, Ana, Radomirović, Mirjana Ž., Ćirković-Veličković, Tanja, "Expression, purification and immunological characterization of recombinant nucleocapsid protein fragment from SARS-CoV-2" in Virology journal, 557 (2021):15-22,
https://doi.org/10.1016/j.virol.2021.01.004 . .

TY  - JOUR
AU  - Đukić, Teodora
AU  - Mladenović, Maja
AU  - Stanić-Vučinić, Dragana
AU  - Radosavljević, Jelena
AU  - Smiljanić, Katarina
AU  - Sabljić, Ljiljana
AU  - Dević, Marija
AU  - Ćujić, Danica R.
AU  - Vasović, Tamara
AU  - Simović, Ana
AU  - Radomirović, Mirjana Ž.
AU  - Ćirković-Veličković, Tanja
PY  - 2021
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/4331
AB  - Serological testing is important method for diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Nucleocapsid (N) protein is the most abundant virus derived protein and strong immunogen. We aimed to find its efficient, low-cost production.SARS-CoV-2 recombinant fragment of nucleocapsid protein (rfNP; 58–419 aa) was expressed in E. coli in soluble form, purified and characterized biochemically and immunologically.Purified rfNP has secondary structure of full-length recombinant N protein, with high percentage of disordered structure (34.2%) and of β-sheet (40.7%). rfNP was tested in immunoblot using sera of COVID-19 convalescent patients. ELISA was optimized with sera of RT-PCR confirmed positive symptomatic patients and healthy individuals. IgG detection sensitivity was 96% (47/50) and specificity 97% (67/68), while IgM detection was slightly lower (94% and 96.5%, respectively).Cost-effective approach for soluble recombinant N protein fragment production was developed, with reliable IgG and IgM antibodies detection of SARS-CoV-2 infection.
PB  - Elsevier
T2  - Virology journal
T1  - Expression, purification and immunological characterization of recombinant nucleocapsid protein fragment from SARS-CoV-2
VL  - 557
SP  - 15
EP  - 22
DO  - 10.1016/j.virol.2021.01.004
ER  - 

@article{
author = "Đukić, Teodora and Mladenović, Maja and Stanić-Vučinić, Dragana and Radosavljević, Jelena and Smiljanić, Katarina and Sabljić, Ljiljana and Dević, Marija and Ćujić, Danica R. and Vasović, Tamara and Simović, Ana and Radomirović, Mirjana Ž. and Ćirković-Veličković, Tanja",
year = "2021",
abstract = "Serological testing is important method for diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Nucleocapsid (N) protein is the most abundant virus derived protein and strong immunogen. We aimed to find its efficient, low-cost production.SARS-CoV-2 recombinant fragment of nucleocapsid protein (rfNP; 58–419 aa) was expressed in E. coli in soluble form, purified and characterized biochemically and immunologically.Purified rfNP has secondary structure of full-length recombinant N protein, with high percentage of disordered structure (34.2%) and of β-sheet (40.7%). rfNP was tested in immunoblot using sera of COVID-19 convalescent patients. ELISA was optimized with sera of RT-PCR confirmed positive symptomatic patients and healthy individuals. IgG detection sensitivity was 96% (47/50) and specificity 97% (67/68), while IgM detection was slightly lower (94% and 96.5%, respectively).Cost-effective approach for soluble recombinant N protein fragment production was developed, with reliable IgG and IgM antibodies detection of SARS-CoV-2 infection.",
publisher = "Elsevier",
journal = "Virology journal",
title = "Expression, purification and immunological characterization of recombinant nucleocapsid protein fragment from SARS-CoV-2",
volume = "557",
pages = "15-22",
doi = "10.1016/j.virol.2021.01.004"
}

Đukić, T., Mladenović, M., Stanić-Vučinić, D., Radosavljević, J., Smiljanić, K., Sabljić, L., Dević, M., Ćujić, D. R., Vasović, T., Simović, A., Radomirović, M. Ž.,& Ćirković-Veličković, T.. (2021). Expression, purification and immunological characterization of recombinant nucleocapsid protein fragment from SARS-CoV-2. in Virology journal
Elsevier., 557, 15-22.
https://doi.org/10.1016/j.virol.2021.01.004

Đukić T, Mladenović M, Stanić-Vučinić D, Radosavljević J, Smiljanić K, Sabljić L, Dević M, Ćujić DR, Vasović T, Simović A, Radomirović MŽ, Ćirković-Veličković T. Expression, purification and immunological characterization of recombinant nucleocapsid protein fragment from SARS-CoV-2. in Virology journal. 2021;557:15-22.
doi:10.1016/j.virol.2021.01.004 .

Đukić, Teodora, Mladenović, Maja, Stanić-Vučinić, Dragana, Radosavljević, Jelena, Smiljanić, Katarina, Sabljić, Ljiljana, Dević, Marija, Ćujić, Danica R., Vasović, Tamara, Simović, Ana, Radomirović, Mirjana Ž., Ćirković-Veličković, Tanja, "Expression, purification and immunological characterization of recombinant nucleocapsid protein fragment from SARS-CoV-2" in Virology journal, 557 (2021):15-22,
https://doi.org/10.1016/j.virol.2021.01.004 . .

TY  - JOUR
AU  - Šunderić, Miloš
AU  - Vasović, Tamara
AU  - Milčić, Miloš K.
AU  - Miljević, Čedo
AU  - Nedić, Olgica
AU  - Nikolić, Milan
AU  - Gligorijević, Nikola
PY  - 2021
UR  - https://www.sciencedirect.com/science/article/pii/S0141813021009284
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/4538
AB  - In this study, the interaction between clozapine, an atypical antipsychotic drug, and alpha-2-macroglobulin (α2M), a multipurpose anti-proteinase, was investigated under simulated (patho) physiological conditions using multiple spectroscopic techniques and molecular modeling. It was found that α2M binds clozapine with a moderate affinity (the binding constant of 0.9 × 105 M−1 at 37 °C). The preferable binding site for both clozapine's atropisomers was revealed to be a large pocket at the interface of C and D monomer subunits of the protein. Hydrogen bonds and the hydrophobic effect were proposed as dominant forces in complex formation. The binding of clozapine did not induce significant conformational change of the protein, as confirmed by virtually unaltered α2M secondary structure and anti-proteinase activity. However, both clozapine and α2M shielded each other from the deleterious influence of strong oxidants: sodium hypochlorite and 2,2′-azobis-2-methyl-propanimidamide dihydrochloride (AAPH). Moreover, clozapine in a concentration range that is usually targeted in the plasma during patients' treatment effectively protected the anti-proteinase activity of α2M under AAPH-induced free radical overproduction. Our results suggest that the cooperation between α2M and clozapine may be a path by which these two molecules synergistically protect neural tissue against injury caused by disturbed proteostasis or oxidative stress.
PB  - Elsevier
T2  - International Journal of Biological Macromolecules
T1  - Antipsychotic clozapine binding to alpha-2-macroglobulin protects interacting partners against oxidation and preserves the anti-proteinase activity of the protein
VL  - 183
SP  - 502
EP  - 512
DO  - 10.1016/j.ijbiomac.2021.04.155
ER  - 

@article{
author = "Šunderić, Miloš and Vasović, Tamara and Milčić, Miloš K. and Miljević, Čedo and Nedić, Olgica and Nikolić, Milan and Gligorijević, Nikola",
year = "2021",
abstract = "In this study, the interaction between clozapine, an atypical antipsychotic drug, and alpha-2-macroglobulin (α2M), a multipurpose anti-proteinase, was investigated under simulated (patho) physiological conditions using multiple spectroscopic techniques and molecular modeling. It was found that α2M binds clozapine with a moderate affinity (the binding constant of 0.9 × 105 M−1 at 37 °C). The preferable binding site for both clozapine's atropisomers was revealed to be a large pocket at the interface of C and D monomer subunits of the protein. Hydrogen bonds and the hydrophobic effect were proposed as dominant forces in complex formation. The binding of clozapine did not induce significant conformational change of the protein, as confirmed by virtually unaltered α2M secondary structure and anti-proteinase activity. However, both clozapine and α2M shielded each other from the deleterious influence of strong oxidants: sodium hypochlorite and 2,2′-azobis-2-methyl-propanimidamide dihydrochloride (AAPH). Moreover, clozapine in a concentration range that is usually targeted in the plasma during patients' treatment effectively protected the anti-proteinase activity of α2M under AAPH-induced free radical overproduction. Our results suggest that the cooperation between α2M and clozapine may be a path by which these two molecules synergistically protect neural tissue against injury caused by disturbed proteostasis or oxidative stress.",
publisher = "Elsevier",
journal = "International Journal of Biological Macromolecules",
title = "Antipsychotic clozapine binding to alpha-2-macroglobulin protects interacting partners against oxidation and preserves the anti-proteinase activity of the protein",
volume = "183",
pages = "502-512",
doi = "10.1016/j.ijbiomac.2021.04.155"
}

Šunderić, M., Vasović, T., Milčić, M. K., Miljević, Č., Nedić, O., Nikolić, M.,& Gligorijević, N.. (2021). Antipsychotic clozapine binding to alpha-2-macroglobulin protects interacting partners against oxidation and preserves the anti-proteinase activity of the protein. in International Journal of Biological Macromolecules
Elsevier., 183, 502-512.
https://doi.org/10.1016/j.ijbiomac.2021.04.155

Šunderić M, Vasović T, Milčić MK, Miljević Č, Nedić O, Nikolić M, Gligorijević N. Antipsychotic clozapine binding to alpha-2-macroglobulin protects interacting partners against oxidation and preserves the anti-proteinase activity of the protein. in International Journal of Biological Macromolecules. 2021;183:502-512.
doi:10.1016/j.ijbiomac.2021.04.155 .

Šunderić, Miloš, Vasović, Tamara, Milčić, Miloš K., Miljević, Čedo, Nedić, Olgica, Nikolić, Milan, Gligorijević, Nikola, "Antipsychotic clozapine binding to alpha-2-macroglobulin protects interacting partners against oxidation and preserves the anti-proteinase activity of the protein" in International Journal of Biological Macromolecules, 183 (2021):502-512,
https://doi.org/10.1016/j.ijbiomac.2021.04.155 . .

TY  - DATA
AU  - Šunderić, Miloš
AU  - Vasović, Tamara
AU  - Milčić, Miloš K.
AU  - Miljević, Čedo
AU  - Nedić, Olgica
AU  - Nikolić, Milan
AU  - Gligorijević, Nikola
PY  - 2021
UR  - https://www.sciencedirect.com/science/article/pii/S0141813021009284
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/4541
PB  - Elsevier
T2  - International Journal of Biological Macromolecules
T1  - Supplementary data for the article: Šunderić, M.; Vasović, T.; Milčić, M.; Miljević, Č.; Nedić, O.; Nikolić, M. R.; Gligorijević, N. Antipsychotic Clozapine Binding to Alpha-2-Macroglobulin Protects Interacting Partners against Oxidation and Preserves the Anti-Proteinase Activity of the Protein. International Journal of Biological Macromolecules 2021, 183, 502–512. https://doi.org/10.1016/j.ijbiomac.2021.04.155.
UR  - https://hdl.handle.net/21.15107/rcub_cherry_4541
ER  - 

@misc{
author = "Šunderić, Miloš and Vasović, Tamara and Milčić, Miloš K. and Miljević, Čedo and Nedić, Olgica and Nikolić, Milan and Gligorijević, Nikola",
year = "2021",
publisher = "Elsevier",
journal = "International Journal of Biological Macromolecules",
title = "Supplementary data for the article: Šunderić, M.; Vasović, T.; Milčić, M.; Miljević, Č.; Nedić, O.; Nikolić, M. R.; Gligorijević, N. Antipsychotic Clozapine Binding to Alpha-2-Macroglobulin Protects Interacting Partners against Oxidation and Preserves the Anti-Proteinase Activity of the Protein. International Journal of Biological Macromolecules 2021, 183, 502–512. https://doi.org/10.1016/j.ijbiomac.2021.04.155.",
url = "https://hdl.handle.net/21.15107/rcub_cherry_4541"
}

Šunderić, M., Vasović, T., Milčić, M. K., Miljević, Č., Nedić, O., Nikolić, M.,& Gligorijević, N.. (2021). Supplementary data for the article: Šunderić, M.; Vasović, T.; Milčić, M.; Miljević, Č.; Nedić, O.; Nikolić, M. R.; Gligorijević, N. Antipsychotic Clozapine Binding to Alpha-2-Macroglobulin Protects Interacting Partners against Oxidation and Preserves the Anti-Proteinase Activity of the Protein. International Journal of Biological Macromolecules 2021, 183, 502–512. https://doi.org/10.1016/j.ijbiomac.2021.04.155.. in International Journal of Biological Macromolecules
Elsevier..
https://hdl.handle.net/21.15107/rcub_cherry_4541

Šunderić M, Vasović T, Milčić MK, Miljević Č, Nedić O, Nikolić M, Gligorijević N. Supplementary data for the article: Šunderić, M.; Vasović, T.; Milčić, M.; Miljević, Č.; Nedić, O.; Nikolić, M. R.; Gligorijević, N. Antipsychotic Clozapine Binding to Alpha-2-Macroglobulin Protects Interacting Partners against Oxidation and Preserves the Anti-Proteinase Activity of the Protein. International Journal of Biological Macromolecules 2021, 183, 502–512. https://doi.org/10.1016/j.ijbiomac.2021.04.155.. in International Journal of Biological Macromolecules. 2021;.
https://hdl.handle.net/21.15107/rcub_cherry_4541 .

Šunderić, Miloš, Vasović, Tamara, Milčić, Miloš K., Miljević, Čedo, Nedić, Olgica, Nikolić, Milan, Gligorijević, Nikola, "Supplementary data for the article: Šunderić, M.; Vasović, T.; Milčić, M.; Miljević, Č.; Nedić, O.; Nikolić, M. R.; Gligorijević, N. Antipsychotic Clozapine Binding to Alpha-2-Macroglobulin Protects Interacting Partners against Oxidation and Preserves the Anti-Proteinase Activity of the Protein. International Journal of Biological Macromolecules 2021, 183, 502–512. https://doi.org/10.1016/j.ijbiomac.2021.04.155." in International Journal of Biological Macromolecules (2021),
https://hdl.handle.net/21.15107/rcub_cherry_4541 .

TY  - CONF
AU  - Vasović, Tamara
AU  - Gligorijević, Nikola
AU  - Lević, Steva M.
AU  - Miljević, Čedo
AU  - Nedić, Olgica
AU  - Nikolić, Milan
PY  - 2021
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/5134
AB  - Clozapine is an atypical antipsychotic used for the treatment of
schizophrenia. Prescribed daily doses of clozapine may reach
over 900 mg/day. Some studies reported a connection between
clozapine usage and the development of thrombosis. Our in vitro
study aimed to provide insight into molecular bases of this observation, investigating clozapine binding to isolated fibrinogen, the
main protein involved in hemostasis. Fibrinogen/clozapine interaction was confirmed by protein fluorescence quenching, with
affinity constant calculated to be 1.7 9 105 M1 and the number
of binding sites more than one. Direct interactions do not affect
the structure of fibrinogen, as determined by UV-VIS spectrometry and Fourier-transform infrared spectroscopy, nor fibrinogen
melting temperature, examined by fluorescence spectroscopy.
However, clozapine binding affected fibrin formation, by reducing coagulation speed and thickness of fibrin fibers. This behavior suggests that in the presence of clozapine, fibrinogen may
acquire thrombogenic characteristics. Although no difference in
fibrin gel porosity was detected, other factors present in the
blood may act synergistically with altered fibrin formation to
modify fibrin clot, thus increasing the risk for development of
thrombosis in individuals on clozapine treatment. By ORAC and
HORAC antioxidant assays, we found that clozapine efficiently
protects fibrinogen from free-radicals oxidation. Since the effect
of clozapine on fibrin formation is dose-dependent, it seems that
the dosage of the medication could be the main factor that determines if clozapine will have a more positive or negative effect on
fibrinogen and coagulation process in vivo.
PB  - FEBS Open Bio © 2021 FEBS
C3  - FEBS Open Bio, virtual 45th FEBS Congress from 3rd to 8th July 2021 (originally planned to be held in Ljubljana, Slovenia)
T1  - Atypical antipsychotic clozapine binds fibrinogen and affects fibrin formation
VL  - 11
SP  - 181
EP  - 181
DO  - doi:10.1002/2211-5463.13205
ER  - 

@conference{
author = "Vasović, Tamara and Gligorijević, Nikola and Lević, Steva M. and Miljević, Čedo and Nedić, Olgica and Nikolić, Milan",
year = "2021",
abstract = "Clozapine is an atypical antipsychotic used for the treatment of
schizophrenia. Prescribed daily doses of clozapine may reach
over 900 mg/day. Some studies reported a connection between
clozapine usage and the development of thrombosis. Our in vitro
study aimed to provide insight into molecular bases of this observation, investigating clozapine binding to isolated fibrinogen, the
main protein involved in hemostasis. Fibrinogen/clozapine interaction was confirmed by protein fluorescence quenching, with
affinity constant calculated to be 1.7 9 105 M1 and the number
of binding sites more than one. Direct interactions do not affect
the structure of fibrinogen, as determined by UV-VIS spectrometry and Fourier-transform infrared spectroscopy, nor fibrinogen
melting temperature, examined by fluorescence spectroscopy.
However, clozapine binding affected fibrin formation, by reducing coagulation speed and thickness of fibrin fibers. This behavior suggests that in the presence of clozapine, fibrinogen may
acquire thrombogenic characteristics. Although no difference in
fibrin gel porosity was detected, other factors present in the
blood may act synergistically with altered fibrin formation to
modify fibrin clot, thus increasing the risk for development of
thrombosis in individuals on clozapine treatment. By ORAC and
HORAC antioxidant assays, we found that clozapine efficiently
protects fibrinogen from free-radicals oxidation. Since the effect
of clozapine on fibrin formation is dose-dependent, it seems that
the dosage of the medication could be the main factor that determines if clozapine will have a more positive or negative effect on
fibrinogen and coagulation process in vivo.",
publisher = "FEBS Open Bio © 2021 FEBS",
journal = "FEBS Open Bio, virtual 45th FEBS Congress from 3rd to 8th July 2021 (originally planned to be held in Ljubljana, Slovenia)",
title = "Atypical antipsychotic clozapine binds fibrinogen and affects fibrin formation",
volume = "11",
pages = "181-181",
doi = "doi:10.1002/2211-5463.13205"
}

Vasović, T., Gligorijević, N., Lević, S. M., Miljević, Č., Nedić, O.,& Nikolić, M.. (2021). Atypical antipsychotic clozapine binds fibrinogen and affects fibrin formation. in FEBS Open Bio, virtual 45th FEBS Congress from 3rd to 8th July 2021 (originally planned to be held in Ljubljana, Slovenia)
FEBS Open Bio © 2021 FEBS., 11, 181-181.
https://doi.org/doi:10.1002/2211-5463.13205

Vasović T, Gligorijević N, Lević SM, Miljević Č, Nedić O, Nikolić M. Atypical antipsychotic clozapine binds fibrinogen and affects fibrin formation. in FEBS Open Bio, virtual 45th FEBS Congress from 3rd to 8th July 2021 (originally planned to be held in Ljubljana, Slovenia). 2021;11:181-181.
doi:doi:10.1002/2211-5463.13205 .

Vasović, Tamara, Gligorijević, Nikola, Lević, Steva M., Miljević, Čedo, Nedić, Olgica, Nikolić, Milan, "Atypical antipsychotic clozapine binds fibrinogen and affects fibrin formation" in FEBS Open Bio, virtual 45th FEBS Congress from 3rd to 8th July 2021 (originally planned to be held in Ljubljana, Slovenia), 11 (2021):181-181,
https://doi.org/doi:10.1002/2211-5463.13205 . .

TY  - CONF
AU  - Smiljanić, Katarina
AU  - Prodić, Ivana
AU  - Đukić, Teodora
AU  - Vasović, Tamara
AU  - Jovanović, Vesna B.
AU  - Ćirković-Veličković, Tanja
PY  - 2021
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/5716
C3  - Proteomics and Metabolomics for Personalized Medicine, XV Italian Proteomics Association Annual Meeting, Catholic University of the Sacred Heart, Roma, Italy, 8th-10th September 2021
T1  - Effects of lysin's and arginige's modifications on trypsin proteolytic efficacy imposed before and after the peanut roasting
SP  - 71
EP  - 71
UR  - https://hdl.handle.net/21.15107/rcub_cherry_5716
ER  - 

@conference{
author = "Smiljanić, Katarina and Prodić, Ivana and Đukić, Teodora and Vasović, Tamara and Jovanović, Vesna B. and Ćirković-Veličković, Tanja",
year = "2021",
journal = "Proteomics and Metabolomics for Personalized Medicine, XV Italian Proteomics Association Annual Meeting, Catholic University of the Sacred Heart, Roma, Italy, 8th-10th September 2021",
title = "Effects of lysin's and arginige's modifications on trypsin proteolytic efficacy imposed before and after the peanut roasting",
pages = "71-71",
url = "https://hdl.handle.net/21.15107/rcub_cherry_5716"
}

Smiljanić, K., Prodić, I., Đukić, T., Vasović, T., Jovanović, V. B.,& Ćirković-Veličković, T.. (2021). Effects of lysin's and arginige's modifications on trypsin proteolytic efficacy imposed before and after the peanut roasting. in Proteomics and Metabolomics for Personalized Medicine, XV Italian Proteomics Association Annual Meeting, Catholic University of the Sacred Heart, Roma, Italy, 8th-10th September 2021, 71-71.
https://hdl.handle.net/21.15107/rcub_cherry_5716

Smiljanić K, Prodić I, Đukić T, Vasović T, Jovanović VB, Ćirković-Veličković T. Effects of lysin's and arginige's modifications on trypsin proteolytic efficacy imposed before and after the peanut roasting. in Proteomics and Metabolomics for Personalized Medicine, XV Italian Proteomics Association Annual Meeting, Catholic University of the Sacred Heart, Roma, Italy, 8th-10th September 2021. 2021;:71-71.
https://hdl.handle.net/21.15107/rcub_cherry_5716 .

Smiljanić, Katarina, Prodić, Ivana, Đukić, Teodora, Vasović, Tamara, Jovanović, Vesna B., Ćirković-Veličković, Tanja, "Effects of lysin's and arginige's modifications on trypsin proteolytic efficacy imposed before and after the peanut roasting" in Proteomics and Metabolomics for Personalized Medicine, XV Italian Proteomics Association Annual Meeting, Catholic University of the Sacred Heart, Roma, Italy, 8th-10th September 2021 (2021):71-71,
https://hdl.handle.net/21.15107/rcub_cherry_5716 .

TY  - CONF
AU  - Đukić, Teodora
AU  - Mladenović, Maja
AU  - Stanić-Vučinić, Dragana
AU  - Radosavljević, Jelena
AU  - Smiljanić, Katarina
AU  - Sabljić, Ljiljana
AU  - Gnjatović, Marija Lj.
AU  - Cujić, Danica
AU  - Vasović, Tamara
AU  - Simović, Ana
AU  - Radomirović, Mirjana Ž.
AU  - Ćirković-Veličković, Tanja
PY  - 2021
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/5735
AB  - Serological testing is important method for diagnosis of severe acute respiratory syndrome coronavirus 2 (SARSCoV-2) infection and for assessment of immunological response after the vaccination. Nucleocapsid (N) protein is the most abundant virus protein and strong immunogen. The aim was develop efficient, low-cost production of N protein large fragment and to characterize it with bottom-up, high-resolution tandem mass spectrometry and immunologically.
SARS-CoV-2 recombinant fragment of nucleocapsid protein (rfNP; 58–419 aa) was expressed in E. coli in soluble form and purified by several chromatographic steps and was subjected to SDS-PAGE and in-gel digested with trypsin.
rfNP was tested in immunoblot using sera of COVID-19 convalescent patients. ELISA was optimized with sera of RT-PCR confirmed positive symptomatic patients and healthy individuals. IgG detection sensitivity was 96% (47/50) and specificity 97% (67/68), while IgM detection was slightly lower. Identity of rfNP was confirmed with high scores and peptide coverage above 80%. Estimation from the value of areas under ion extracted chromatographic curves is that only up to 0,03% of the total band protein quantity belongs to host proteins, while rfNP share is well above 99,9%, resulting in highly pure nucleocapsid protein preparation.
C3  - Proteomics and Metabolomics for Personalized Medicine, XV Italian Proteomics Association Annual Meeting, Catholic University of the Sacred Heart, Roma, Italy, 8th-10th September 2021
T1  - Proteomic and immunological characterization of recombinantly expressed nucleocapsid SARS-CoV 2 protein fragment in E. coli
SP  - 50
EP  - 50
UR  - https://hdl.handle.net/21.15107/rcub_cherry_5735
ER  - 

@conference{
author = "Đukić, Teodora and Mladenović, Maja and Stanić-Vučinić, Dragana and Radosavljević, Jelena and Smiljanić, Katarina and Sabljić, Ljiljana and Gnjatović, Marija Lj. and Cujić, Danica and Vasović, Tamara and Simović, Ana and Radomirović, Mirjana Ž. and Ćirković-Veličković, Tanja",
year = "2021",
abstract = "Serological testing is important method for diagnosis of severe acute respiratory syndrome coronavirus 2 (SARSCoV-2) infection and for assessment of immunological response after the vaccination. Nucleocapsid (N) protein is the most abundant virus protein and strong immunogen. The aim was develop efficient, low-cost production of N protein large fragment and to characterize it with bottom-up, high-resolution tandem mass spectrometry and immunologically.
SARS-CoV-2 recombinant fragment of nucleocapsid protein (rfNP; 58–419 aa) was expressed in E. coli in soluble form and purified by several chromatographic steps and was subjected to SDS-PAGE and in-gel digested with trypsin.
rfNP was tested in immunoblot using sera of COVID-19 convalescent patients. ELISA was optimized with sera of RT-PCR confirmed positive symptomatic patients and healthy individuals. IgG detection sensitivity was 96% (47/50) and specificity 97% (67/68), while IgM detection was slightly lower. Identity of rfNP was confirmed with high scores and peptide coverage above 80%. Estimation from the value of areas under ion extracted chromatographic curves is that only up to 0,03% of the total band protein quantity belongs to host proteins, while rfNP share is well above 99,9%, resulting in highly pure nucleocapsid protein preparation.",
journal = "Proteomics and Metabolomics for Personalized Medicine, XV Italian Proteomics Association Annual Meeting, Catholic University of the Sacred Heart, Roma, Italy, 8th-10th September 2021",
title = "Proteomic and immunological characterization of recombinantly expressed nucleocapsid SARS-CoV 2 protein fragment in E. coli",
pages = "50-50",
url = "https://hdl.handle.net/21.15107/rcub_cherry_5735"
}

Đukić, T., Mladenović, M., Stanić-Vučinić, D., Radosavljević, J., Smiljanić, K., Sabljić, L., Gnjatović, M. Lj., Cujić, D., Vasović, T., Simović, A., Radomirović, M. Ž.,& Ćirković-Veličković, T.. (2021). Proteomic and immunological characterization of recombinantly expressed nucleocapsid SARS-CoV 2 protein fragment in E. coli. in Proteomics and Metabolomics for Personalized Medicine, XV Italian Proteomics Association Annual Meeting, Catholic University of the Sacred Heart, Roma, Italy, 8th-10th September 2021, 50-50.
https://hdl.handle.net/21.15107/rcub_cherry_5735

Đukić T, Mladenović M, Stanić-Vučinić D, Radosavljević J, Smiljanić K, Sabljić L, Gnjatović ML, Cujić D, Vasović T, Simović A, Radomirović MŽ, Ćirković-Veličković T. Proteomic and immunological characterization of recombinantly expressed nucleocapsid SARS-CoV 2 protein fragment in E. coli. in Proteomics and Metabolomics for Personalized Medicine, XV Italian Proteomics Association Annual Meeting, Catholic University of the Sacred Heart, Roma, Italy, 8th-10th September 2021. 2021;:50-50.
https://hdl.handle.net/21.15107/rcub_cherry_5735 .

Đukić, Teodora, Mladenović, Maja, Stanić-Vučinić, Dragana, Radosavljević, Jelena, Smiljanić, Katarina, Sabljić, Ljiljana, Gnjatović, Marija Lj., Cujić, Danica, Vasović, Tamara, Simović, Ana, Radomirović, Mirjana Ž., Ćirković-Veličković, Tanja, "Proteomic and immunological characterization of recombinantly expressed nucleocapsid SARS-CoV 2 protein fragment in E. coli" in Proteomics and Metabolomics for Personalized Medicine, XV Italian Proteomics Association Annual Meeting, Catholic University of the Sacred Heart, Roma, Italy, 8th-10th September 2021 (2021):50-50,
https://hdl.handle.net/21.15107/rcub_cherry_5735 .

TY  - CONF
AU  - Maja, Mladenović
AU  - Đukić, Teodora
AU  - Vasović, Tamara
AU  - Stanić-Vučinić, Dragana
AU  - Smiljanić, Katarina
AU  - Radosavljević, Jelena
AU  - Sabljić, Ljiljana
AU  - Dević, Marija
AU  - Cujić, Danica
AU  - Ćirković-Veličković, Tanja
PY  - 2021
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/5737
AB  - Serological testing is important method for diagnosis of severe acute respiratory syndrome coronavirus 2 (SARSCoV-2) infection. Nucleocapsid (N) protein is the most abundant virus derived protein and strong immunogen. We aimed to find its efficient, low-cost production, suitable for serological diagnosis. SARS-CoV-2 recombinant fragment of nucleocapsid protein (rfNP; 58–419 aa) was expressed in E. coli in soluble form, purified by immobilized metal ion affinity chromatography and strong cation exchange chromatography after which it was analyzed by Mass and CD spectrometry and characterized biochemically and immunologically. Purified rfNP has secondary structure of full-length recombinant N protein, with high percentage of disordered structure (34.2%) and of β-sheet (40.7%). rfNP was tested in immunoblot using sera of COVID-19 convalescent patients. ELISA was optimized with sera of RT-PCR confirmed positive symptomatic patients and healthy individuals. IgG detection sensitivity was 96% (47/50) and specificity 97% (67/68), while IgM detection was
slightly lower (94% and 96.5%, respectively). Cost-effective approach for soluble recombinant N protein fragment production was developed, with reliable IgG and IgM antibodies detection of SARS-CoV-2 infection.
C3  - FoodEnTwin Symposium: Novel analytical approaches in food and environmental sciences, June 16-18, 2021 Belgrade, Serbia
T1  - Expression, purification and immunological characterization of recombinant protein fragment from SARS-CoV-2
SP  - 33
EP  - 33
UR  - https://hdl.handle.net/21.15107/rcub_cherry_5737
ER  - 

@conference{
author = "Maja, Mladenović and Đukić, Teodora and Vasović, Tamara and Stanić-Vučinić, Dragana and Smiljanić, Katarina and Radosavljević, Jelena and Sabljić, Ljiljana and Dević, Marija and Cujić, Danica and Ćirković-Veličković, Tanja",
year = "2021",
abstract = "Serological testing is important method for diagnosis of severe acute respiratory syndrome coronavirus 2 (SARSCoV-2) infection. Nucleocapsid (N) protein is the most abundant virus derived protein and strong immunogen. We aimed to find its efficient, low-cost production, suitable for serological diagnosis. SARS-CoV-2 recombinant fragment of nucleocapsid protein (rfNP; 58–419 aa) was expressed in E. coli in soluble form, purified by immobilized metal ion affinity chromatography and strong cation exchange chromatography after which it was analyzed by Mass and CD spectrometry and characterized biochemically and immunologically. Purified rfNP has secondary structure of full-length recombinant N protein, with high percentage of disordered structure (34.2%) and of β-sheet (40.7%). rfNP was tested in immunoblot using sera of COVID-19 convalescent patients. ELISA was optimized with sera of RT-PCR confirmed positive symptomatic patients and healthy individuals. IgG detection sensitivity was 96% (47/50) and specificity 97% (67/68), while IgM detection was
slightly lower (94% and 96.5%, respectively). Cost-effective approach for soluble recombinant N protein fragment production was developed, with reliable IgG and IgM antibodies detection of SARS-CoV-2 infection.",
journal = "FoodEnTwin Symposium: Novel analytical approaches in food and environmental sciences, June 16-18, 2021 Belgrade, Serbia",
title = "Expression, purification and immunological characterization of recombinant protein fragment from SARS-CoV-2",
pages = "33-33",
url = "https://hdl.handle.net/21.15107/rcub_cherry_5737"
}

Maja, M., Đukić, T., Vasović, T., Stanić-Vučinić, D., Smiljanić, K., Radosavljević, J., Sabljić, L., Dević, M., Cujić, D.,& Ćirković-Veličković, T.. (2021). Expression, purification and immunological characterization of recombinant protein fragment from SARS-CoV-2. in FoodEnTwin Symposium: Novel analytical approaches in food and environmental sciences, June 16-18, 2021 Belgrade, Serbia, 33-33.
https://hdl.handle.net/21.15107/rcub_cherry_5737

Maja M, Đukić T, Vasović T, Stanić-Vučinić D, Smiljanić K, Radosavljević J, Sabljić L, Dević M, Cujić D, Ćirković-Veličković T. Expression, purification and immunological characterization of recombinant protein fragment from SARS-CoV-2. in FoodEnTwin Symposium: Novel analytical approaches in food and environmental sciences, June 16-18, 2021 Belgrade, Serbia. 2021;:33-33.
https://hdl.handle.net/21.15107/rcub_cherry_5737 .

Maja, Mladenović, Đukić, Teodora, Vasović, Tamara, Stanić-Vučinić, Dragana, Smiljanić, Katarina, Radosavljević, Jelena, Sabljić, Ljiljana, Dević, Marija, Cujić, Danica, Ćirković-Veličković, Tanja, "Expression, purification and immunological characterization of recombinant protein fragment from SARS-CoV-2" in FoodEnTwin Symposium: Novel analytical approaches in food and environmental sciences, June 16-18, 2021 Belgrade, Serbia (2021):33-33,
https://hdl.handle.net/21.15107/rcub_cherry_5737 .

TY  - CONF
AU  - Smiljanić, Katarina
AU  - Prodić, Ivana
AU  - Đukić, Teodora
AU  - Vasović, Tamara
AU  - Jovanović, Vesna B.
AU  - Ćirković-Veličković, Tanja
PY  - 2021
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/5773
AB  - Background: We have undertaken study on our porcine-derived trypsin generated
proteomic data of the major peanut allergen Ara h 1 from the raw and roasted peanut, to
assess possible facilitating/hindrance effects on trypsin digestion efficacy caused by post-
translational and chemical modifications (PTMs) positioned on K/R residues. If potential
hindrance effects caused by PTMs are observed with porcine trypsin, then they can be just
augmented and more pronounced within human intestinal digestion. The logic for such
reasoning is in inferior performance of human trypsin compared to porcine-derived used in
proteomic digestion protocols, also the lower trypsin-to-sample ratio and much shorter
digestion times, even though gastric digestion precedes and trypsin is not the sole digestive
enzyme.
Methods: Novel method was developed to decipher outcomes at scissile bonds using PEAKS
Studio-X+ in reassessment of high-resolution tandem mass spectrometry data on 18h-long
trypsin digestion protocol.
Results: In eight modified K/R residues involving methylation, dihydroxy and formylation,
differences in extent of miscleavage between modified and unmodified peptides, were
significantly higher (>10%) in modified peptides.
Conclusion: It is important to elucidate impact of modifications on trypsin digestion
performance, but also on other proteases involved in digestion process due to possible
effects on allergenicity of food proteins/peptides.
PB  - INFOGEST Cost action, INRAE, Teagasc LTD.
C3  - Virtual International Conference on Food Digestion 6th and 7th May, 2021
T1  - Trypsin as a proteomic probe to assess food protein digestibility in relation to post- translational modifications
SP  - 18
EP  - 18
UR  - https://hdl.handle.net/21.15107/rcub_cherry_5773
ER  - 

@conference{
author = "Smiljanić, Katarina and Prodić, Ivana and Đukić, Teodora and Vasović, Tamara and Jovanović, Vesna B. and Ćirković-Veličković, Tanja",
year = "2021",
abstract = "Background: We have undertaken study on our porcine-derived trypsin generated
proteomic data of the major peanut allergen Ara h 1 from the raw and roasted peanut, to
assess possible facilitating/hindrance effects on trypsin digestion efficacy caused by post-
translational and chemical modifications (PTMs) positioned on K/R residues. If potential
hindrance effects caused by PTMs are observed with porcine trypsin, then they can be just
augmented and more pronounced within human intestinal digestion. The logic for such
reasoning is in inferior performance of human trypsin compared to porcine-derived used in
proteomic digestion protocols, also the lower trypsin-to-sample ratio and much shorter
digestion times, even though gastric digestion precedes and trypsin is not the sole digestive
enzyme.
Methods: Novel method was developed to decipher outcomes at scissile bonds using PEAKS
Studio-X+ in reassessment of high-resolution tandem mass spectrometry data on 18h-long
trypsin digestion protocol.
Results: In eight modified K/R residues involving methylation, dihydroxy and formylation,
differences in extent of miscleavage between modified and unmodified peptides, were
significantly higher (>10%) in modified peptides.
Conclusion: It is important to elucidate impact of modifications on trypsin digestion
performance, but also on other proteases involved in digestion process due to possible
effects on allergenicity of food proteins/peptides.",
publisher = "INFOGEST Cost action, INRAE, Teagasc LTD.",
journal = "Virtual International Conference on Food Digestion 6th and 7th May, 2021",
title = "Trypsin as a proteomic probe to assess food protein digestibility in relation to post- translational modifications",
pages = "18-18",
url = "https://hdl.handle.net/21.15107/rcub_cherry_5773"
}

Smiljanić, K., Prodić, I., Đukić, T., Vasović, T., Jovanović, V. B.,& Ćirković-Veličković, T.. (2021). Trypsin as a proteomic probe to assess food protein digestibility in relation to post- translational modifications. in Virtual International Conference on Food Digestion 6th and 7th May, 2021
INFOGEST Cost action, INRAE, Teagasc LTD.., 18-18.
https://hdl.handle.net/21.15107/rcub_cherry_5773

Smiljanić K, Prodić I, Đukić T, Vasović T, Jovanović VB, Ćirković-Veličković T. Trypsin as a proteomic probe to assess food protein digestibility in relation to post- translational modifications. in Virtual International Conference on Food Digestion 6th and 7th May, 2021. 2021;:18-18.
https://hdl.handle.net/21.15107/rcub_cherry_5773 .

Smiljanić, Katarina, Prodić, Ivana, Đukić, Teodora, Vasović, Tamara, Jovanović, Vesna B., Ćirković-Veličković, Tanja, "Trypsin as a proteomic probe to assess food protein digestibility in relation to post- translational modifications" in Virtual International Conference on Food Digestion 6th and 7th May, 2021 (2021):18-18,
https://hdl.handle.net/21.15107/rcub_cherry_5773 .