Stojanović, Marijana M.

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Authority KeyName Variants
orcid::0000-0002-8204-4183
  • Stojanović, Marijana M. (11)
  • Stojanović, Marijana (4)
  • Stojanovic, M. (2)
Projects
Allergens, antibodies, enzymes and small physiologically important molecules: design, structure, function and relevance CAPSIDO – Developement of the assays for detection of SARS Cov-2 virus capsid proteins in biological fluids of COVID19 patients
Ministry of Education, Science and Technological Development, Republic of Serbia, Grant no. 451-03-68/2020-14/200168 (University of Belgrade, Faculty of Chemistry) Ministry of Education, Science and Technological Development, Republic of Serbia, Grant no. 451-03-68/2020-14/200177 (Immunology Research Centre 'Branislav Janković' Torlak, Belgrade)
Ispitivanje strukture i funkcije biološki važnih makromolekula u fiziološkim i patološkim stanjima Study of structure-function relationships in the plant cell wall and modifications of the wall structure by enzyme engineering
Characterization and application of fungal metabolites and assessment of new biofungicides potential Synthesis, processing and characterization of nanostructured materials for application in the field of energy, mechanical engineering, environmental protection and biomedicine
Serbian Academy of Sciences and Arts GA No. F-26. Ghent University Global Campus and Belgian Special Research Fund BOF StG No. 01N01718.
The pathogenetic mechanism in hematological malignancies Ministry of Education, Science and Technological Development, Republic of Serbia, Grant no. 451-03-68/2020-14/200007 (University of Belgrade, Institute for Biological Research 'Siniša Stanković')
Ministry of Education, Science and Technological Development, Republic of Serbia, Grant no. 451-03-68/2020-14/200026 (University of Belgrade, Institute of Chemistry, Technology and Metallurgy - IChTM) Ministry of Education, Science and Technological Development, Republic of Serbia, Grant no. 451-03-68/2020-14/200288 (Innovation Center of the Faculty of Chemistry)
This work was institutionally funded by the Institute for Specific Prophylaxis and Tropical Medicine, Medical University of Vienna, Austria, and partially supported by the Austrian Research Promotion Agency (FFG project number 822768). Превенција и одговор на COVID-19 у угроженим подручјима - одржива производња серолошког IgG теста за SARS CoV-2 у Србији - LVP-BPA UNDP 00121484/2020-02

Author's Bibliography

Sandwich ELISA for the Quantification of Nucleocapsid Protein of SARS-CoV-2 Based on Polyclonal Antibodies from Two Different Species

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

(MDPI, 2023)

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 . .

Expression of recombinant SARS-CoV-2 nucleocapsid protein in mammalian cells

Sibinčić, Nikolina; Krstić-Ristivojević, Maja; Stojanović, Marijana; Mladenović Stokanić, Maja; Vasović, Tamara; Ćirković-Veličković, Tanja; Stojadinović, Marija

(Beograde : Serbian Biochemical Society, 2023)

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 .

Dobijanje rekombinantnog imunogenog fragmenta proteina nukleokapsida SARS-CoV-2 virusa za proizvodnju reagenasa i dijagnostičkih testova na novi korona virus

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

(2023)

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 .

SARS CoV-2 nucleocapsid-based diagnostic tests and serological response in allergic children

Ćirković-Veličković, Tanja; Radomirović, Mirjana Ž.; Simović, Ana; Jovanović, Vesna B.; Ćujić, Danica R.; Gnjatović, Marija Lj.; Stojanović, Marijana

(2022)

TY  - CONF
AU  - Ćirković-Veličković, Tanja
AU  - Radomirović, Mirjana Ž.
AU  - Simović, Ana
AU  - Jovanović, Vesna B.
AU  - Ćujić, Danica R.
AU  - Gnjatović, Marija Lj.
AU  - Stojanović, Marijana
PY  - 2022
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/5777
AB  - RT-PCR remains the primary method of diagnosing SARS-CoV-2. Serological diagnosis of COVID-19 is simple and does not require complex techniques and equipment, rendering it suitable for rapid detection and massive screening. However, serological tests cannot confirm SARS-CoV-2, and results will be false negative when antibody concentrations fall below detection limit. Serological testing is important method for diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS CoV-2) infection. Antigen-based tests show high specificity, but sensitivity is relatively low. For development and further improvement of both serological and antigen-based tests, availability of recombinantly producted SARS CoV-2 antigens is needed and during the periods of high demend in the pandemic, it was a limited factor both for research and diagnostic tests production by national producers of tests. 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 beta-sheet (40.7%). rfNP was tested in immunoblot using sera of COVID-19 convalescent patients. Cost-effective approach for soluble recombinant N protein fragment production was developed, with reliable IgG and IgM antibodies detection of SARS-CoV-2 infection. 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). Application of serological ELISA for seropositivity detection in 200 allergic children in comparison to non-allergic did not show a bias towards any of the tested groups.  Furthermore, we have optimized production of rfNP at a large scale to raise specific antibodies to N-protein in rabbit and mice and develop an in house ELISA for quantification of N protein in biological fluids. Test was clinically validated in 200 PCR positive patients and showed a sensitivity of around 50%.
C3  - International Congress on Molecular Immunology and Allergology (IMAC-2022), Moscow, December 1-3, 2022
T1  - SARS CoV-2 nucleocapsid-based diagnostic tests and serological response in allergic children
UR  - https://hdl.handle.net/21.15107/rcub_cherry_5777
ER  - 
@conference{
author = "Ćirković-Veličković, Tanja and Radomirović, Mirjana Ž. and Simović, Ana and Jovanović, Vesna B. and Ćujić, Danica R. and Gnjatović, Marija Lj. and Stojanović, Marijana",
year = "2022",
abstract = "RT-PCR remains the primary method of diagnosing SARS-CoV-2. Serological diagnosis of COVID-19 is simple and does not require complex techniques and equipment, rendering it suitable for rapid detection and massive screening. However, serological tests cannot confirm SARS-CoV-2, and results will be false negative when antibody concentrations fall below detection limit. Serological testing is important method for diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS CoV-2) infection. Antigen-based tests show high specificity, but sensitivity is relatively low. For development and further improvement of both serological and antigen-based tests, availability of recombinantly producted SARS CoV-2 antigens is needed and during the periods of high demend in the pandemic, it was a limited factor both for research and diagnostic tests production by national producers of tests. 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 beta-sheet (40.7%). rfNP was tested in immunoblot using sera of COVID-19 convalescent patients. Cost-effective approach for soluble recombinant N protein fragment production was developed, with reliable IgG and IgM antibodies detection of SARS-CoV-2 infection. 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). Application of serological ELISA for seropositivity detection in 200 allergic children in comparison to non-allergic did not show a bias towards any of the tested groups.  Furthermore, we have optimized production of rfNP at a large scale to raise specific antibodies to N-protein in rabbit and mice and develop an in house ELISA for quantification of N protein in biological fluids. Test was clinically validated in 200 PCR positive patients and showed a sensitivity of around 50%.",
journal = "International Congress on Molecular Immunology and Allergology (IMAC-2022), Moscow, December 1-3, 2022",
title = "SARS CoV-2 nucleocapsid-based diagnostic tests and serological response in allergic children",
url = "https://hdl.handle.net/21.15107/rcub_cherry_5777"
}
Ćirković-Veličković, T., Radomirović, M. Ž., Simović, A., Jovanović, V. B., Ćujić, D. R., Gnjatović, M. Lj.,& Stojanović, M.. (2022). SARS CoV-2 nucleocapsid-based diagnostic tests and serological response in allergic children. in International Congress on Molecular Immunology and Allergology (IMAC-2022), Moscow, December 1-3, 2022.
https://hdl.handle.net/21.15107/rcub_cherry_5777
Ćirković-Veličković T, Radomirović MŽ, Simović A, Jovanović VB, Ćujić DR, Gnjatović ML, Stojanović M. SARS CoV-2 nucleocapsid-based diagnostic tests and serological response in allergic children. in International Congress on Molecular Immunology and Allergology (IMAC-2022), Moscow, December 1-3, 2022. 2022;.
https://hdl.handle.net/21.15107/rcub_cherry_5777 .
Ćirković-Veličković, Tanja, Radomirović, Mirjana Ž., Simović, Ana, Jovanović, Vesna B., Ćujić, Danica R., Gnjatović, Marija Lj., Stojanović, Marijana, "SARS CoV-2 nucleocapsid-based diagnostic tests and serological response in allergic children" in International Congress on Molecular Immunology and Allergology (IMAC-2022), Moscow, December 1-3, 2022 (2022),
https://hdl.handle.net/21.15107/rcub_cherry_5777 .

Razvoj sendvič ELISA testa specifičnog za SARS-CoV-2 N-protein

Radomirović, Mirjana Ž.; Simović, Ana; Udovički, Božidar D.; Krstić-Ristivojević, Maja; Sabljić, Ljiljana Z.; Lukić, Ivana D.; Glamočlija, Sofija Đ.; Ćujić, Danica R.; Gnjatović, Marija Lj.; Stojanović, Marijana M.; Stanić-Vučinić, Dragana; Radosavljević, Jelena; Ćirković-Veličković, Tanja

(Belgrade : Serbian Chemical Society, 2022)

TY  - CONF
AU  - Radomirović, Mirjana Ž.
AU  - Simović, Ana
AU  - Udovički, Božidar D.
AU  - Krstić-Ristivojević, Maja
AU  - Sabljić, Ljiljana Z.
AU  - Lukić, Ivana D.
AU  - Glamočlija, Sofija Đ.
AU  - Ćujić, Danica R.
AU  - Gnjatović, Marija Lj.
AU  - Stojanović, Marijana M.
AU  - Stanić-Vučinić, Dragana
AU  - Radosavljević, Jelena
AU  - Ćirković-Veličković, Tanja
PY  - 2022
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/5361
AB  - Ta na dijagnoza ljudi sa sumnjom na infekciju SARS-CoV-2 je od suštinskog zna aja za
suzbijanje globalnog širenja COVID-19. Prisustvo SARS-CoV-2 može se otkriti RT-PCRom (otkriva RNK virusa) ili detekcijom prisustva virusnih antigena u biološkim te nostima
ELISA-om ili sli nom tehnikom koje koriste antitela razvijena u životinjama. Cilj studije
je bio uspostavljanje kvantitativnog testa koji se zasniva na koriš enju poliklonskih seruma
za rutinsko odre ivanje koncentracije SARS-CoV-2 nukleokapsidnog proteina merenjam
apsorbancije u standardnoj mikrotitarskoj plo ici sa 96 bunara. Za potrebe razvoja testa
proizveden je rekombinantni N-protein i koriš en za proizvodnju antiseruma u miševima i
ze evima. Proizvedeni antiserumi su pre iš eni i odre en im je titar. Poliklonskiantiserumi
visokog afiniteta specifi ni za N-protein koriš eni su za razvoj ELISA testa specifi nog za
ovaj protein. Test se zasniva na koriš enju poliklonskih seruma miševa koji su adherirani
na dno bunara mikrotitarske plo ice za hvatanje N-proteina iz uzorka. Razli ite
koncentracije rekombinantnog N-proteina su koriš ene za standardnu krivu za
kvantifikaciju proteina. N-protein vezan za antitela miševa je detektovan ze jim
poliklonskim serumom i anti-ze jim antitelom povezanim sa enzimom koji obezbe uje
spektrofotometrijsko merenje. Uspešno smo razvili prototip ELISA testa za kvantifikaciju
N-proteina sa granicom detekcije u opsegu od ng/mL. Prose na vrednost LOD za prototip
ELISA testa za detekciju N-proteina je 9,2 ng/mL, dok je prose na vrednost LOQ
10,2 ng/mL. Pokazali smo da su proizvedeni poliklonski antiserumi pogodni za detekciju
N-proteina sa sli nim ili boljim afinitetom i specifi noš u od komercijalnih antitela.
Štaviše, prototip ELISA testa se može koristiti sa zadovoljavaju om pouzdanoš u za
kvantifikaciju N-proteina u uzorcima bogatim proteinima, poput ljudskih seruma.
AB  - The accurate diagnosis of people with suspected infection with the SARS-CoV-2 is
essential to curb the global spread of COVID-19. The presence of SARS-CoV-2 can be
detected by RT-PCR (it detects RNA of the virus) or by the presence of viral antigens in
biological fluids in ELISA or similar techniques using antibodies developed in animals.
The aim of the study was the establishment of a quantitative polyclonal sera-based test for
routine measurement of the concentration of SARS CoV-2 nucleocapsid protein using
absorbance measurement in a standard 96-well microtiter plate. For the purposes of the test
development, recombinant N protein was produced and used for the production of mice
and rabbit antisera. Produced antisera were purified and titer was determined. High-affinity
polyclonal N-protein specific antisera were used for N-protein specific ELISA test
development. The test is based on mice polyclonal sera adhered to microtiter plate bottom
for the capture of the N protein from the specimen. Various concentrations of the
recombinant N-protein were used to generate a standard curve for protein quantification.
The N-protein bound to the mice antibodies was detected with rabbit polyclonal sera and
anti-rabbit antibody coupled to an enzyme that provides spectrophotometric measurement.
We have successfully developed the prototype ELISA for the quantification of N-protein
with the detection limit being in the range of ng/mL. The average LOD value for the
prototype ELISA was determined to be 9.2 ng/mL, while the average LOQ value was
10.2 ng/mL. We have demonstrated that produced polyclonal antisera are suitable for the
detection of N-protein with affinity and specificity similar to, or better than commercial
antibodies. Furthermore, the prototype ELISA can be used with satisfactory confidence for
quantification of the N-protein in protein-rich samples, similar to human sera.
PB  - Belgrade : Serbian Chemical Society
C3  - 58th Meeting of the Serbian Chemical Society, Belgrade, Serbia, 9th-10th June, 2022. In: Book of Abstracts and Proceedings
T1  - Razvoj sendvič ELISA testa specifičnog za SARS-CoV-2 N-protein
T1  - Development of SARS-CoV-2 N-protein specific capture ELISA
SP  - 65
EP  - 66
UR  - https://hdl.handle.net/21.15107/rcub_cherry_5361
ER  - 
@conference{
author = "Radomirović, Mirjana Ž. and Simović, Ana and Udovički, Božidar D. and Krstić-Ristivojević, Maja and Sabljić, Ljiljana Z. and Lukić, Ivana D. and Glamočlija, Sofija Đ. and Ćujić, Danica R. and Gnjatović, Marija Lj. and Stojanović, Marijana M. and Stanić-Vučinić, Dragana and Radosavljević, Jelena and Ćirković-Veličković, Tanja",
year = "2022",
abstract = "Ta na dijagnoza ljudi sa sumnjom na infekciju SARS-CoV-2 je od suštinskog zna aja za
suzbijanje globalnog širenja COVID-19. Prisustvo SARS-CoV-2 može se otkriti RT-PCRom (otkriva RNK virusa) ili detekcijom prisustva virusnih antigena u biološkim te nostima
ELISA-om ili sli nom tehnikom koje koriste antitela razvijena u životinjama. Cilj studije
je bio uspostavljanje kvantitativnog testa koji se zasniva na koriš enju poliklonskih seruma
za rutinsko odre ivanje koncentracije SARS-CoV-2 nukleokapsidnog proteina merenjam
apsorbancije u standardnoj mikrotitarskoj plo ici sa 96 bunara. Za potrebe razvoja testa
proizveden je rekombinantni N-protein i koriš en za proizvodnju antiseruma u miševima i
ze evima. Proizvedeni antiserumi su pre iš eni i odre en im je titar. Poliklonskiantiserumi
visokog afiniteta specifi ni za N-protein koriš eni su za razvoj ELISA testa specifi nog za
ovaj protein. Test se zasniva na koriš enju poliklonskih seruma miševa koji su adherirani
na dno bunara mikrotitarske plo ice za hvatanje N-proteina iz uzorka. Razli ite
koncentracije rekombinantnog N-proteina su koriš ene za standardnu krivu za
kvantifikaciju proteina. N-protein vezan za antitela miševa je detektovan ze jim
poliklonskim serumom i anti-ze jim antitelom povezanim sa enzimom koji obezbe uje
spektrofotometrijsko merenje. Uspešno smo razvili prototip ELISA testa za kvantifikaciju
N-proteina sa granicom detekcije u opsegu od ng/mL. Prose na vrednost LOD za prototip
ELISA testa za detekciju N-proteina je 9,2 ng/mL, dok je prose na vrednost LOQ
10,2 ng/mL. Pokazali smo da su proizvedeni poliklonski antiserumi pogodni za detekciju
N-proteina sa sli nim ili boljim afinitetom i specifi noš u od komercijalnih antitela.
Štaviše, prototip ELISA testa se može koristiti sa zadovoljavaju om pouzdanoš u za
kvantifikaciju N-proteina u uzorcima bogatim proteinima, poput ljudskih seruma., The accurate diagnosis of people with suspected infection with the SARS-CoV-2 is
essential to curb the global spread of COVID-19. The presence of SARS-CoV-2 can be
detected by RT-PCR (it detects RNA of the virus) or by the presence of viral antigens in
biological fluids in ELISA or similar techniques using antibodies developed in animals.
The aim of the study was the establishment of a quantitative polyclonal sera-based test for
routine measurement of the concentration of SARS CoV-2 nucleocapsid protein using
absorbance measurement in a standard 96-well microtiter plate. For the purposes of the test
development, recombinant N protein was produced and used for the production of mice
and rabbit antisera. Produced antisera were purified and titer was determined. High-affinity
polyclonal N-protein specific antisera were used for N-protein specific ELISA test
development. The test is based on mice polyclonal sera adhered to microtiter plate bottom
for the capture of the N protein from the specimen. Various concentrations of the
recombinant N-protein were used to generate a standard curve for protein quantification.
The N-protein bound to the mice antibodies was detected with rabbit polyclonal sera and
anti-rabbit antibody coupled to an enzyme that provides spectrophotometric measurement.
We have successfully developed the prototype ELISA for the quantification of N-protein
with the detection limit being in the range of ng/mL. The average LOD value for the
prototype ELISA was determined to be 9.2 ng/mL, while the average LOQ value was
10.2 ng/mL. We have demonstrated that produced polyclonal antisera are suitable for the
detection of N-protein with affinity and specificity similar to, or better than commercial
antibodies. Furthermore, the prototype ELISA can be used with satisfactory confidence for
quantification of the N-protein in protein-rich samples, similar to human sera.",
publisher = "Belgrade : Serbian Chemical Society",
journal = "58th Meeting of the Serbian Chemical Society, Belgrade, Serbia, 9th-10th June, 2022. In: Book of Abstracts and Proceedings",
title = "Razvoj sendvič ELISA testa specifičnog za SARS-CoV-2 N-protein, Development of SARS-CoV-2 N-protein specific capture ELISA",
pages = "65-66",
url = "https://hdl.handle.net/21.15107/rcub_cherry_5361"
}
Radomirović, M. Ž., Simović, A., Udovički, B. D., Krstić-Ristivojević, M., Sabljić, L. Z., Lukić, I. D., Glamočlija, S. Đ., Ćujić, D. R., Gnjatović, M. Lj., Stojanović, M. M., Stanić-Vučinić, D., Radosavljević, J.,& Ćirković-Veličković, T.. (2022). Razvoj sendvič ELISA testa specifičnog za SARS-CoV-2 N-protein. in 58th Meeting of the Serbian Chemical Society, Belgrade, Serbia, 9th-10th June, 2022. In: Book of Abstracts and Proceedings
Belgrade : Serbian Chemical Society., 65-66.
https://hdl.handle.net/21.15107/rcub_cherry_5361
Radomirović MŽ, Simović A, Udovički BD, Krstić-Ristivojević M, Sabljić LZ, Lukić ID, Glamočlija SĐ, Ćujić DR, Gnjatović ML, Stojanović MM, Stanić-Vučinić D, Radosavljević J, Ćirković-Veličković T. Razvoj sendvič ELISA testa specifičnog za SARS-CoV-2 N-protein. in 58th Meeting of the Serbian Chemical Society, Belgrade, Serbia, 9th-10th June, 2022. In: Book of Abstracts and Proceedings. 2022;:65-66.
https://hdl.handle.net/21.15107/rcub_cherry_5361 .
Radomirović, Mirjana Ž., Simović, Ana, Udovički, Božidar D., Krstić-Ristivojević, Maja, Sabljić, Ljiljana Z., Lukić, Ivana D., Glamočlija, Sofija Đ., Ćujić, Danica R., Gnjatović, Marija Lj., Stojanović, Marijana M., Stanić-Vučinić, Dragana, Radosavljević, Jelena, Ćirković-Veličković, Tanja, "Razvoj sendvič ELISA testa specifičnog za SARS-CoV-2 N-protein" in 58th Meeting of the Serbian Chemical Society, Belgrade, Serbia, 9th-10th June, 2022. In: Book of Abstracts and Proceedings (2022):65-66,
https://hdl.handle.net/21.15107/rcub_cherry_5361 .

Razvoj sendvič ELISA testa specifičnog za SARS-CoV-2 N-protein

Radomirović, Mirjana Ž.; Simović, Ana; Udovički, Božidar D.; Krstić-Ristivojević, Maja; Sabljić, Ljiljana Z.; Lukić, Ivana D.; Glamočlija, Sofija Đ.; Ćujić, Danica R.; Gnjatović, Marija Lj.; Stojanović, Marijana M.; Stanić-Vučinić, Dragana; Radosavljević, Jelena; Ćirković-Veličković, Tanja

(Belgrade : Serbian Chemical Society, 2022)

TY  - CONF
AU  - Radomirović, Mirjana Ž.
AU  - Simović, Ana
AU  - Udovički, Božidar D.
AU  - Krstić-Ristivojević, Maja
AU  - Sabljić, Ljiljana Z.
AU  - Lukić, Ivana D.
AU  - Glamočlija, Sofija Đ.
AU  - Ćujić, Danica R.
AU  - Gnjatović, Marija Lj.
AU  - Stojanović, Marijana M.
AU  - Stanić-Vučinić, Dragana
AU  - Radosavljević, Jelena
AU  - Ćirković-Veličković, Tanja
PY  - 2022
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/5362
AB  - Ta na dijagnoza ljudi sa sumnjom na infekciju SARS-CoV-2 je od suštinskog zna aja zasuzbijanje globalnog širenja COVID-19. Prisustvo SARS-CoV-2 može se otkriti RT-PCRom (otkriva RNK virusa) ili detekcijom prisustva virusnih antigena u biološkim te nostimaELISA-om ili sli nom tehnikom koje koriste antitela razvijena u životinjama. Cilj studijeje bio uspostavljanje kvantitativnog testa koji se zasniva na koriš enju poliklonskih serumaza rutinsko odre ivanje koncentracije SARS-CoV-2 nukleokapsidnog proteina merenjamapsorbancije u standardnoj mikrotitarskoj plo ici sa 96 bunara. Za potrebe razvoja testaproizveden je rekombinantni N-protein i koriš en za proizvodnju antiseruma u miševima ize evima. Proizvedeni antiserumi su pre iš eni i odre en im je titar. Poliklonskiantiserumivisokog afiniteta specifi ni za N-protein koriš eni su za razvoj ELISA testa specifi nog zaovaj protein. Test se zasniva na koriš enju poliklonskih seruma miševa koji su adheriranina dno bunara mikrotitarske plo ice za hvatanje N-proteina iz uzorka. Razli itekoncentracije rekombinantnog N-proteina su koriš ene za standardnu krivu zakvantifikaciju proteina. N-protein vezan za antitela miševa je detektovan ze jimpoliklonskim serumom i anti-ze jim antitelom povezanim sa enzimom koji obezbe ujespektrofotometrijsko merenje. Uspešno smo razvili prototip ELISA testa za kvantifikacijuN-proteina sa granicom detekcije u opsegu od ng/mL. Prose na vrednost LOD za prototipELISA testa za detekciju N-proteina je 9,2 ng/mL, dok je prose na vrednost LOQ10,2 ng/mL. Pokazali smo da su proizvedeni poliklonski antiserumi pogodni za detekcijuN-proteina sa sli nim ili boljim afinitetom i specifi noš u od komercijalnih antitela.Štaviše, prototip ELISA testa se može koristiti sa zadovoljavaju om pouzdanoš u zakvantifikaciju N-proteina u uzorcima bogatim proteinima, poput ljudskih seruma.
AB  - The accurate diagnosis of people with suspected infection with the SARS-CoV-2 isessential to curb the global spread of COVID-19. The presence of SARS-CoV-2 can bedetected by RT-PCR (it detects RNA of the virus) or by the presence of viral antigens inbiological fluids in ELISA or similar techniques using antibodies developed in animals.The aim of the study was the establishment of a quantitative polyclonal sera-based test forroutine measurement of the concentration of SARS CoV-2 nucleocapsid protein usingabsorbance measurement in a standard 96-well microtiter plate. For the purposes of the testdevelopment, recombinant N protein was produced and used for the production of miceand rabbit antisera. Produced antisera were purified and titer was determined. High-affinitypolyclonal N-protein specific antisera were used for N-protein specific ELISA testdevelopment. The test is based on mice polyclonal sera adhered to microtiter plate bottomfor the capture of the N protein from the specimen. Various concentrations of therecombinant N-protein were used to generate a standard curve for protein quantification.The N-protein bound to the mice antibodies was detected with rabbit polyclonal sera andanti-rabbit antibody coupled to an enzyme that provides spectrophotometric measurement.We have successfully developed the prototype ELISA for the quantification of N-proteinwith the detection limit being in the range of ng/mL. The average LOD value for theprototype ELISA was determined to be 9.2 ng/mL, while the average LOQ value was10.2 ng/mL. We have demonstrated that produced polyclonal antisera are suitable for thedetection of N-protein with affinity and specificity similar to, or better than commercialantibodies. Furthermore, the prototype ELISA can be used with satisfactory confidence forquantification of the N-protein in protein-rich samples, similar to human sera.
PB  - Belgrade : Serbian Chemical Society
C3  - 58th Meeting of the Serbian Chemical Society, Belgrade, Serbia, 9th-10th June, 2022. In: Book of Abstracts and Proceedings
T1  - Razvoj sendvič ELISA testa specifičnog za SARS-CoV-2 N-protein
T1  - Development of SARS-CoV-2 N-protein specific capture ELISA
UR  - https://hdl.handle.net/21.15107/rcub_cherry_5362
ER  - 
@conference{
author = "Radomirović, Mirjana Ž. and Simović, Ana and Udovički, Božidar D. and Krstić-Ristivojević, Maja and Sabljić, Ljiljana Z. and Lukić, Ivana D. and Glamočlija, Sofija Đ. and Ćujić, Danica R. and Gnjatović, Marija Lj. and Stojanović, Marijana M. and Stanić-Vučinić, Dragana and Radosavljević, Jelena and Ćirković-Veličković, Tanja",
year = "2022",
abstract = "Ta na dijagnoza ljudi sa sumnjom na infekciju SARS-CoV-2 je od suštinskog zna aja zasuzbijanje globalnog širenja COVID-19. Prisustvo SARS-CoV-2 može se otkriti RT-PCRom (otkriva RNK virusa) ili detekcijom prisustva virusnih antigena u biološkim te nostimaELISA-om ili sli nom tehnikom koje koriste antitela razvijena u životinjama. Cilj studijeje bio uspostavljanje kvantitativnog testa koji se zasniva na koriš enju poliklonskih serumaza rutinsko odre ivanje koncentracije SARS-CoV-2 nukleokapsidnog proteina merenjamapsorbancije u standardnoj mikrotitarskoj plo ici sa 96 bunara. Za potrebe razvoja testaproizveden je rekombinantni N-protein i koriš en za proizvodnju antiseruma u miševima ize evima. Proizvedeni antiserumi su pre iš eni i odre en im je titar. Poliklonskiantiserumivisokog afiniteta specifi ni za N-protein koriš eni su za razvoj ELISA testa specifi nog zaovaj protein. Test se zasniva na koriš enju poliklonskih seruma miševa koji su adheriranina dno bunara mikrotitarske plo ice za hvatanje N-proteina iz uzorka. Razli itekoncentracije rekombinantnog N-proteina su koriš ene za standardnu krivu zakvantifikaciju proteina. N-protein vezan za antitela miševa je detektovan ze jimpoliklonskim serumom i anti-ze jim antitelom povezanim sa enzimom koji obezbe ujespektrofotometrijsko merenje. Uspešno smo razvili prototip ELISA testa za kvantifikacijuN-proteina sa granicom detekcije u opsegu od ng/mL. Prose na vrednost LOD za prototipELISA testa za detekciju N-proteina je 9,2 ng/mL, dok je prose na vrednost LOQ10,2 ng/mL. Pokazali smo da su proizvedeni poliklonski antiserumi pogodni za detekcijuN-proteina sa sli nim ili boljim afinitetom i specifi noš u od komercijalnih antitela.Štaviše, prototip ELISA testa se može koristiti sa zadovoljavaju om pouzdanoš u zakvantifikaciju N-proteina u uzorcima bogatim proteinima, poput ljudskih seruma., The accurate diagnosis of people with suspected infection with the SARS-CoV-2 isessential to curb the global spread of COVID-19. The presence of SARS-CoV-2 can bedetected by RT-PCR (it detects RNA of the virus) or by the presence of viral antigens inbiological fluids in ELISA or similar techniques using antibodies developed in animals.The aim of the study was the establishment of a quantitative polyclonal sera-based test forroutine measurement of the concentration of SARS CoV-2 nucleocapsid protein usingabsorbance measurement in a standard 96-well microtiter plate. For the purposes of the testdevelopment, recombinant N protein was produced and used for the production of miceand rabbit antisera. Produced antisera were purified and titer was determined. High-affinitypolyclonal N-protein specific antisera were used for N-protein specific ELISA testdevelopment. The test is based on mice polyclonal sera adhered to microtiter plate bottomfor the capture of the N protein from the specimen. Various concentrations of therecombinant N-protein were used to generate a standard curve for protein quantification.The N-protein bound to the mice antibodies was detected with rabbit polyclonal sera andanti-rabbit antibody coupled to an enzyme that provides spectrophotometric measurement.We have successfully developed the prototype ELISA for the quantification of N-proteinwith the detection limit being in the range of ng/mL. The average LOD value for theprototype ELISA was determined to be 9.2 ng/mL, while the average LOQ value was10.2 ng/mL. We have demonstrated that produced polyclonal antisera are suitable for thedetection of N-protein with affinity and specificity similar to, or better than commercialantibodies. Furthermore, the prototype ELISA can be used with satisfactory confidence forquantification of the N-protein in protein-rich samples, similar to human sera.",
publisher = "Belgrade : Serbian Chemical Society",
journal = "58th Meeting of the Serbian Chemical Society, Belgrade, Serbia, 9th-10th June, 2022. In: Book of Abstracts and Proceedings",
title = "Razvoj sendvič ELISA testa specifičnog za SARS-CoV-2 N-protein, Development of SARS-CoV-2 N-protein specific capture ELISA",
url = "https://hdl.handle.net/21.15107/rcub_cherry_5362"
}
Radomirović, M. Ž., Simović, A., Udovički, B. D., Krstić-Ristivojević, M., Sabljić, L. Z., Lukić, I. D., Glamočlija, S. Đ., Ćujić, D. R., Gnjatović, M. Lj., Stojanović, M. M., Stanić-Vučinić, D., Radosavljević, J.,& Ćirković-Veličković, T.. (2022). Razvoj sendvič ELISA testa specifičnog za SARS-CoV-2 N-protein. in 58th Meeting of the Serbian Chemical Society, Belgrade, Serbia, 9th-10th June, 2022. In: Book of Abstracts and Proceedings
Belgrade : Serbian Chemical Society..
https://hdl.handle.net/21.15107/rcub_cherry_5362
Radomirović MŽ, Simović A, Udovički BD, Krstić-Ristivojević M, Sabljić LZ, Lukić ID, Glamočlija SĐ, Ćujić DR, Gnjatović ML, Stojanović MM, Stanić-Vučinić D, Radosavljević J, Ćirković-Veličković T. Razvoj sendvič ELISA testa specifičnog za SARS-CoV-2 N-protein. in 58th Meeting of the Serbian Chemical Society, Belgrade, Serbia, 9th-10th June, 2022. In: Book of Abstracts and Proceedings. 2022;.
https://hdl.handle.net/21.15107/rcub_cherry_5362 .
Radomirović, Mirjana Ž., Simović, Ana, Udovički, Božidar D., Krstić-Ristivojević, Maja, Sabljić, Ljiljana Z., Lukić, Ivana D., Glamočlija, Sofija Đ., Ćujić, Danica R., Gnjatović, Marija Lj., Stojanović, Marijana M., Stanić-Vučinić, Dragana, Radosavljević, Jelena, Ćirković-Veličković, Tanja, "Razvoj sendvič ELISA testa specifičnog za SARS-CoV-2 N-protein" in 58th Meeting of the Serbian Chemical Society, Belgrade, Serbia, 9th-10th June, 2022. In: Book of Abstracts and Proceedings (2022),
https://hdl.handle.net/21.15107/rcub_cherry_5362 .

Recombinant Bet v 1-BanLec chimera modulates functional characteristics of peritoneal murine macrophages by promoting IL-10 secretion

Protić-Rosić, Isidora; Nešić, Andrijana N.; Lukić, Ivana; Miljković, Radmila; Popović, Dragan M.; Atanasković-Marković, Marina; Stojanović, Marijana M.; Gavrović-Jankulović, Marija

(Elsevier, 2021)

TY  - JOUR
AU  - Protić-Rosić, Isidora
AU  - Nešić, Andrijana N.
AU  - Lukić, Ivana
AU  - Miljković, Radmila
AU  - Popović, Dragan M.
AU  - Atanasković-Marković, Marina
AU  - Stojanović, Marijana M.
AU  - Gavrović-Jankulović, Marija
PY  - 2021
UR  - https://www.sciencedirect.com/science/article/pii/S0161589021001905
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/4604
AB  - Allergen-specific immunotherapy (AIT) is a desensitizing treatment for allergic diseases that corrects the underlined pathological immune response to innocuous protein antigens, called allergens. Recombinant allergens employed in the AIT allowed the production of well-defined formulations that possessed consistent quality but were often less efficient than natural allergen extracts. Combining recombinant allergens with an adjuvant or immunomodulatory agent could improve AIT efficacy. This study aimed to perform structural and functional characterization of newly designed recombinant chimera composed of the Bet v 1, the major birch pollen allergen, and Banana Lectin (BanLec), TLR2, and CD14 binding protein, for the application in AIT. rBet v 1-BanLec chimera was designed in silico and expressed as a soluble fraction in Escherichia coli. Purified rBet v 1-BanLec (33.4 kDa) retained BanLec-associated biological activity of carbohydrate-binding and preserved IgE reactive epitopes of Bet v 1. The chimera revealed secondary structures with predominant β sheets. The immunomodulatory capacity of rBet v 1-BanLec tested on macrophages showed changes in myeloperoxidase activity, reduced NO production, and significant alterations in the production of cytokines when compared to both rBanLec and rBet v 1. Comparing to rBet v 1, rBet v 1-BanLec was demonstrated to be more efficient promoter of IL-10 production as well as weaker inducer of NO production and secretion of pro-inflammatory cytokines TNFα, and IL-6. The ability of rBet v 1-BanLec to promote IL-10 in together with the preserved 3D structure of Bet v 1 part implies that the construct might exert a beneficial effect in the allergen-specific immunotherapy.
PB  - Elsevier
T2  - Molecular Immunology
T2  - Molecular ImmunologyMolecular Immunology
T1  - Recombinant Bet v 1-BanLec chimera modulates functional characteristics of peritoneal murine macrophages by promoting IL-10 secretion
VL  - 138
SP  - 58
EP  - 67
DO  - 10.1016/j.molimm.2021.06.015
ER  - 
@article{
author = "Protić-Rosić, Isidora and Nešić, Andrijana N. and Lukić, Ivana and Miljković, Radmila and Popović, Dragan M. and Atanasković-Marković, Marina and Stojanović, Marijana M. and Gavrović-Jankulović, Marija",
year = "2021",
abstract = "Allergen-specific immunotherapy (AIT) is a desensitizing treatment for allergic diseases that corrects the underlined pathological immune response to innocuous protein antigens, called allergens. Recombinant allergens employed in the AIT allowed the production of well-defined formulations that possessed consistent quality but were often less efficient than natural allergen extracts. Combining recombinant allergens with an adjuvant or immunomodulatory agent could improve AIT efficacy. This study aimed to perform structural and functional characterization of newly designed recombinant chimera composed of the Bet v 1, the major birch pollen allergen, and Banana Lectin (BanLec), TLR2, and CD14 binding protein, for the application in AIT. rBet v 1-BanLec chimera was designed in silico and expressed as a soluble fraction in Escherichia coli. Purified rBet v 1-BanLec (33.4 kDa) retained BanLec-associated biological activity of carbohydrate-binding and preserved IgE reactive epitopes of Bet v 1. The chimera revealed secondary structures with predominant β sheets. The immunomodulatory capacity of rBet v 1-BanLec tested on macrophages showed changes in myeloperoxidase activity, reduced NO production, and significant alterations in the production of cytokines when compared to both rBanLec and rBet v 1. Comparing to rBet v 1, rBet v 1-BanLec was demonstrated to be more efficient promoter of IL-10 production as well as weaker inducer of NO production and secretion of pro-inflammatory cytokines TNFα, and IL-6. The ability of rBet v 1-BanLec to promote IL-10 in together with the preserved 3D structure of Bet v 1 part implies that the construct might exert a beneficial effect in the allergen-specific immunotherapy.",
publisher = "Elsevier",
journal = "Molecular Immunology, Molecular ImmunologyMolecular Immunology",
title = "Recombinant Bet v 1-BanLec chimera modulates functional characteristics of peritoneal murine macrophages by promoting IL-10 secretion",
volume = "138",
pages = "58-67",
doi = "10.1016/j.molimm.2021.06.015"
}
Protić-Rosić, I., Nešić, A. N., Lukić, I., Miljković, R., Popović, D. M., Atanasković-Marković, M., Stojanović, M. M.,& Gavrović-Jankulović, M.. (2021). Recombinant Bet v 1-BanLec chimera modulates functional characteristics of peritoneal murine macrophages by promoting IL-10 secretion. in Molecular Immunology
Elsevier., 138, 58-67.
https://doi.org/10.1016/j.molimm.2021.06.015
Protić-Rosić I, Nešić AN, Lukić I, Miljković R, Popović DM, Atanasković-Marković M, Stojanović MM, Gavrović-Jankulović M. Recombinant Bet v 1-BanLec chimera modulates functional characteristics of peritoneal murine macrophages by promoting IL-10 secretion. in Molecular Immunology. 2021;138:58-67.
doi:10.1016/j.molimm.2021.06.015 .
Protić-Rosić, Isidora, Nešić, Andrijana N., Lukić, Ivana, Miljković, Radmila, Popović, Dragan M., Atanasković-Marković, Marina, Stojanović, Marijana M., Gavrović-Jankulović, Marija, "Recombinant Bet v 1-BanLec chimera modulates functional characteristics of peritoneal murine macrophages by promoting IL-10 secretion" in Molecular Immunology, 138 (2021):58-67,
https://doi.org/10.1016/j.molimm.2021.06.015 . .
1
1

Cross-Reactive Effects of Vaccines: Heterologous Immunity between Tetanus and Chlamydia

Stojanović, Marijana M.; Lukić, Ivana; Marinković, Emilija; Kovačević, Ana; Miljković, Radmila; Tobias, Joshua; Schabussova, Irma; Zlatović, Mario; Barisani-Asenbauer, Talin; Wiedermann, Ursula; Inic-Kanada, Aleksandra

(2020)

TY  - JOUR
AU  - Stojanović, Marijana M.
AU  - Lukić, Ivana
AU  - Marinković, Emilija
AU  - Kovačević, Ana
AU  - Miljković, Radmila
AU  - Tobias, Joshua
AU  - Schabussova, Irma
AU  - Zlatović, Mario
AU  - Barisani-Asenbauer, Talin
AU  - Wiedermann, Ursula
AU  - Inic-Kanada, Aleksandra
PY  - 2020
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/4292
AB  - Vaccines can have heterologous effects on the immune system, i.e., effects other than triggering an immune response against the disease targeted by the vaccine. We investigated whether monoclonal antibodies (mAbs) specific for tetanus could cross-react with Chlamydia and confer heterologous protection against chlamydial infection. The capability of two tetanus-specific mAbs, namely mAb26 and mAb51, to prevent chlamydial infection has been assessed: (i) in vitro, by performing a neutralization assay using human conjunctival epithelial (HCjE) cells infected with Chlamydia trachomatis serovar B, and (ii) in vivo, by using a guinea pig model of Chlamydiacaviae-induced inclusion conjunctivitis. The mAb26 has been superior in comparison with mAb51 in the prevention of chlamydial infection in HCjE cells. The mAb26 has conferred &asymp;40% inhibition of the infection, compared to less than 5% inhibition in the presence of the mAb51. In vivo, mAb26 significantly diminished ocular pathology intensity in guinea pigs infected with C. caviae compared to either the mAb51-treated or sham-treated guinea pigs. Our data provide insights that tetanus immunization generates antibodies which induce heterologous chlamydial immunity and promote protection beyond the intended target pathogen.
T2  - Vaccines
T1  - Cross-Reactive Effects of Vaccines: Heterologous Immunity between Tetanus and Chlamydia
VL  - 8
IS  - 4
SP  - 719
DO  - 10.3390/vaccines8040719
ER  - 
@article{
author = "Stojanović, Marijana M. and Lukić, Ivana and Marinković, Emilija and Kovačević, Ana and Miljković, Radmila and Tobias, Joshua and Schabussova, Irma and Zlatović, Mario and Barisani-Asenbauer, Talin and Wiedermann, Ursula and Inic-Kanada, Aleksandra",
year = "2020",
abstract = "Vaccines can have heterologous effects on the immune system, i.e., effects other than triggering an immune response against the disease targeted by the vaccine. We investigated whether monoclonal antibodies (mAbs) specific for tetanus could cross-react with Chlamydia and confer heterologous protection against chlamydial infection. The capability of two tetanus-specific mAbs, namely mAb26 and mAb51, to prevent chlamydial infection has been assessed: (i) in vitro, by performing a neutralization assay using human conjunctival epithelial (HCjE) cells infected with Chlamydia trachomatis serovar B, and (ii) in vivo, by using a guinea pig model of Chlamydiacaviae-induced inclusion conjunctivitis. The mAb26 has been superior in comparison with mAb51 in the prevention of chlamydial infection in HCjE cells. The mAb26 has conferred &asymp;40% inhibition of the infection, compared to less than 5% inhibition in the presence of the mAb51. In vivo, mAb26 significantly diminished ocular pathology intensity in guinea pigs infected with C. caviae compared to either the mAb51-treated or sham-treated guinea pigs. Our data provide insights that tetanus immunization generates antibodies which induce heterologous chlamydial immunity and promote protection beyond the intended target pathogen.",
journal = "Vaccines",
title = "Cross-Reactive Effects of Vaccines: Heterologous Immunity between Tetanus and Chlamydia",
volume = "8",
number = "4",
pages = "719",
doi = "10.3390/vaccines8040719"
}
Stojanović, M. M., Lukić, I., Marinković, E., Kovačević, A., Miljković, R., Tobias, J., Schabussova, I., Zlatović, M., Barisani-Asenbauer, T., Wiedermann, U.,& Inic-Kanada, A.. (2020). Cross-Reactive Effects of Vaccines: Heterologous Immunity between Tetanus and Chlamydia. in Vaccines, 8(4), 719.
https://doi.org/10.3390/vaccines8040719
Stojanović MM, Lukić I, Marinković E, Kovačević A, Miljković R, Tobias J, Schabussova I, Zlatović M, Barisani-Asenbauer T, Wiedermann U, Inic-Kanada A. Cross-Reactive Effects of Vaccines: Heterologous Immunity between Tetanus and Chlamydia. in Vaccines. 2020;8(4):719.
doi:10.3390/vaccines8040719 .
Stojanović, Marijana M., Lukić, Ivana, Marinković, Emilija, Kovačević, Ana, Miljković, Radmila, Tobias, Joshua, Schabussova, Irma, Zlatović, Mario, Barisani-Asenbauer, Talin, Wiedermann, Ursula, Inic-Kanada, Aleksandra, "Cross-Reactive Effects of Vaccines: Heterologous Immunity between Tetanus and Chlamydia" in Vaccines, 8, no. 4 (2020):719,
https://doi.org/10.3390/vaccines8040719 . .
2
5
2
4
3

Supplementary data for article: Marinkovic, E.; Djokic, R.; Lukic, I.; Filipovic, A.; Inic-Kanada, A.; Kosanovic, D.; Gavrovic-Jankulovic, M.; Stojanovic, M. Modulation of Functional Characteristics of Resident and Thioglycollate-Elicited Peritoneal Murine Macrophages by a Recombinant Banana Lectin. PLoS ONE 2017, 12 (2). https://doi.org/10.1371/journal.pone.017246

Marinković, Emilija; Đokić, Radmila; Lukić, Ivana; Filipović, Ana; Inić-Kanada, Aleksandra; Kosanović, Dejana; Gavrović-Jankulović, Marija; Stojanović, Marijana M.

(Public Library Science, San Francisco, 2017)

TY  - DATA
AU  - Marinković, Emilija
AU  - Đokić, Radmila
AU  - Lukić, Ivana
AU  - Filipović, Ana
AU  - Inić-Kanada, Aleksandra
AU  - Kosanović, Dejana
AU  - Gavrović-Jankulović, Marija
AU  - Stojanović, Marijana M.
PY  - 2017
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/3037
PB  - Public Library Science, San Francisco
T2  - PLoS ONE
T1  - Supplementary data for article: Marinkovic, E.; Djokic, R.; Lukic, I.; Filipovic, A.; Inic-Kanada, A.; Kosanovic, D.; Gavrovic-Jankulovic, M.; Stojanovic, M. Modulation of Functional Characteristics of Resident and Thioglycollate-Elicited Peritoneal Murine Macrophages by a Recombinant Banana Lectin. PLoS ONE 2017, 12 (2). https://doi.org/10.1371/journal.pone.017246
UR  - https://hdl.handle.net/21.15107/rcub_cherry_3037
ER  - 
@misc{
author = "Marinković, Emilija and Đokić, Radmila and Lukić, Ivana and Filipović, Ana and Inić-Kanada, Aleksandra and Kosanović, Dejana and Gavrović-Jankulović, Marija and Stojanović, Marijana M.",
year = "2017",
publisher = "Public Library Science, San Francisco",
journal = "PLoS ONE",
title = "Supplementary data for article: Marinkovic, E.; Djokic, R.; Lukic, I.; Filipovic, A.; Inic-Kanada, A.; Kosanovic, D.; Gavrovic-Jankulovic, M.; Stojanovic, M. Modulation of Functional Characteristics of Resident and Thioglycollate-Elicited Peritoneal Murine Macrophages by a Recombinant Banana Lectin. PLoS ONE 2017, 12 (2). https://doi.org/10.1371/journal.pone.017246",
url = "https://hdl.handle.net/21.15107/rcub_cherry_3037"
}
Marinković, E., Đokić, R., Lukić, I., Filipović, A., Inić-Kanada, A., Kosanović, D., Gavrović-Jankulović, M.,& Stojanović, M. M.. (2017). Supplementary data for article: Marinkovic, E.; Djokic, R.; Lukic, I.; Filipovic, A.; Inic-Kanada, A.; Kosanovic, D.; Gavrovic-Jankulovic, M.; Stojanovic, M. Modulation of Functional Characteristics of Resident and Thioglycollate-Elicited Peritoneal Murine Macrophages by a Recombinant Banana Lectin. PLoS ONE 2017, 12 (2). https://doi.org/10.1371/journal.pone.017246. in PLoS ONE
Public Library Science, San Francisco..
https://hdl.handle.net/21.15107/rcub_cherry_3037
Marinković E, Đokić R, Lukić I, Filipović A, Inić-Kanada A, Kosanović D, Gavrović-Jankulović M, Stojanović MM. Supplementary data for article: Marinkovic, E.; Djokic, R.; Lukic, I.; Filipovic, A.; Inic-Kanada, A.; Kosanovic, D.; Gavrovic-Jankulovic, M.; Stojanovic, M. Modulation of Functional Characteristics of Resident and Thioglycollate-Elicited Peritoneal Murine Macrophages by a Recombinant Banana Lectin. PLoS ONE 2017, 12 (2). https://doi.org/10.1371/journal.pone.017246. in PLoS ONE. 2017;.
https://hdl.handle.net/21.15107/rcub_cherry_3037 .
Marinković, Emilija, Đokić, Radmila, Lukić, Ivana, Filipović, Ana, Inić-Kanada, Aleksandra, Kosanović, Dejana, Gavrović-Jankulović, Marija, Stojanović, Marijana M., "Supplementary data for article: Marinkovic, E.; Djokic, R.; Lukic, I.; Filipovic, A.; Inic-Kanada, A.; Kosanovic, D.; Gavrovic-Jankulovic, M.; Stojanovic, M. Modulation of Functional Characteristics of Resident and Thioglycollate-Elicited Peritoneal Murine Macrophages by a Recombinant Banana Lectin. PLoS ONE 2017, 12 (2). https://doi.org/10.1371/journal.pone.017246" in PLoS ONE (2017),
https://hdl.handle.net/21.15107/rcub_cherry_3037 .

Modulation of functional characteristics of resident and thioglycollate-elicited peritoneal murine macrophages by a recombinant banana lectin

Marinković, Emilija; Đokić, Radmila; Lukić, Ivana; Filipović, Ana; Inić-Kanada, Aleksandra; Kosanović, Dejana; Gavrović-Jankulović, Marija; Stojanović, Marijana M.

(Public Library Science, San Francisco, 2017)

TY  - JOUR
AU  - Marinković, Emilija
AU  - Đokić, Radmila
AU  - Lukić, Ivana
AU  - Filipović, Ana
AU  - Inić-Kanada, Aleksandra
AU  - Kosanović, Dejana
AU  - Gavrović-Jankulović, Marija
AU  - Stojanović, Marijana M.
PY  - 2017
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/2417
AB  - We demonstrated that a recombinant banana lectin (rBanLec), which structural characteristics and physiological impacts highly resemble those reported for its natural counterparts, binds murine peritoneal macrophages and specifically modulates their functional characteristics. By using rBanLec in concentrations ranging from 1 mu g to 10 mu g to stimulate resident (RMs) and thioglycollate-elicited (TGMs) peritoneal macrophages from BALB/c and C57BL/6 mice, we have shown that effects of rBanLec stimulation depend on its concentration but also on the functional status of macrophages and their genetic background. rBanLec, in a positive dose-dependent manner, promotes the proliferation of TGMs from both BALB/c and C57BL/6 mice, while its mitogenic influence on RMs is significantly lower (BALB/c mice) or not detectable (C57BL/6 mice). In all peritoneal macrophages, irrespective of their type and genetic background, rBanLec, in a positive dose dependent manner, enhances the secretion of IL-10. rBanLec stimulation of RMs from both BALB/c and C57BL/6 resulted in a positive dose-dependent promotion of proinflammatory phenotype (enhancement of NO production and IL-12 and TNF alpha secretion, reduction of arginase activity). Positive dose-dependent skewing toward proinflammatory phenotype was also observed in TGMs from C57BL/6 mice. However, the enhancement of rBanLec stimulation promotes skewing of TGMs from BALB/c mice towards anti-inflammatory profile (reduction of NO production and IL-12 secretion, enhancement of arginase activity and TGF alpha and IL-4 secretion). Moreover, we established that rBanLec binds oligosaccharide structures of TLR2 and CD14 and that blocking of signaling via these receptors significantly impairs the production of TNFa and NO in BALB/c macrophages. Since the outcome of rBanLec stimulation depends on rBanLec concentration as well as on the functional characteristics of its target cells and their genetic background, further studies are needed to investigate its effects under physiological and specific pathological conditions.
PB  - Public Library Science, San Francisco
T2  - PLoS One / Public Library of Science
T1  - Modulation of functional characteristics of resident and thioglycollate-elicited peritoneal murine macrophages by a recombinant banana lectin
VL  - 12
IS  - 2
DO  - 10.1371/journal.pone.0172469
ER  - 
@article{
author = "Marinković, Emilija and Đokić, Radmila and Lukić, Ivana and Filipović, Ana and Inić-Kanada, Aleksandra and Kosanović, Dejana and Gavrović-Jankulović, Marija and Stojanović, Marijana M.",
year = "2017",
abstract = "We demonstrated that a recombinant banana lectin (rBanLec), which structural characteristics and physiological impacts highly resemble those reported for its natural counterparts, binds murine peritoneal macrophages and specifically modulates their functional characteristics. By using rBanLec in concentrations ranging from 1 mu g to 10 mu g to stimulate resident (RMs) and thioglycollate-elicited (TGMs) peritoneal macrophages from BALB/c and C57BL/6 mice, we have shown that effects of rBanLec stimulation depend on its concentration but also on the functional status of macrophages and their genetic background. rBanLec, in a positive dose-dependent manner, promotes the proliferation of TGMs from both BALB/c and C57BL/6 mice, while its mitogenic influence on RMs is significantly lower (BALB/c mice) or not detectable (C57BL/6 mice). In all peritoneal macrophages, irrespective of their type and genetic background, rBanLec, in a positive dose dependent manner, enhances the secretion of IL-10. rBanLec stimulation of RMs from both BALB/c and C57BL/6 resulted in a positive dose-dependent promotion of proinflammatory phenotype (enhancement of NO production and IL-12 and TNF alpha secretion, reduction of arginase activity). Positive dose-dependent skewing toward proinflammatory phenotype was also observed in TGMs from C57BL/6 mice. However, the enhancement of rBanLec stimulation promotes skewing of TGMs from BALB/c mice towards anti-inflammatory profile (reduction of NO production and IL-12 secretion, enhancement of arginase activity and TGF alpha and IL-4 secretion). Moreover, we established that rBanLec binds oligosaccharide structures of TLR2 and CD14 and that blocking of signaling via these receptors significantly impairs the production of TNFa and NO in BALB/c macrophages. Since the outcome of rBanLec stimulation depends on rBanLec concentration as well as on the functional characteristics of its target cells and their genetic background, further studies are needed to investigate its effects under physiological and specific pathological conditions.",
publisher = "Public Library Science, San Francisco",
journal = "PLoS One / Public Library of Science",
title = "Modulation of functional characteristics of resident and thioglycollate-elicited peritoneal murine macrophages by a recombinant banana lectin",
volume = "12",
number = "2",
doi = "10.1371/journal.pone.0172469"
}
Marinković, E., Đokić, R., Lukić, I., Filipović, A., Inić-Kanada, A., Kosanović, D., Gavrović-Jankulović, M.,& Stojanović, M. M.. (2017). Modulation of functional characteristics of resident and thioglycollate-elicited peritoneal murine macrophages by a recombinant banana lectin. in PLoS One / Public Library of Science
Public Library Science, San Francisco., 12(2).
https://doi.org/10.1371/journal.pone.0172469
Marinković E, Đokić R, Lukić I, Filipović A, Inić-Kanada A, Kosanović D, Gavrović-Jankulović M, Stojanović MM. Modulation of functional characteristics of resident and thioglycollate-elicited peritoneal murine macrophages by a recombinant banana lectin. in PLoS One / Public Library of Science. 2017;12(2).
doi:10.1371/journal.pone.0172469 .
Marinković, Emilija, Đokić, Radmila, Lukić, Ivana, Filipović, Ana, Inić-Kanada, Aleksandra, Kosanović, Dejana, Gavrović-Jankulović, Marija, Stojanović, Marijana M., "Modulation of functional characteristics of resident and thioglycollate-elicited peritoneal murine macrophages by a recombinant banana lectin" in PLoS One / Public Library of Science, 12, no. 2 (2017),
https://doi.org/10.1371/journal.pone.0172469 . .
8
6
6
5

Lignin model compound in alginate hydrogel: a strong antimicrobial agent with high potential in wound treatment

Spasojević, Dragica; Zmejkoski, Danica; Glamočlija, Jasmina; Nikolić, Miloš; Soković, Marina; Milošević, Verica; Jarić, Ivana; Stojanović, Marijana M.; Marinković, Emilija; Barisani-Asenbauer, Talin; Prodanović, Radivoje; Jovanović, Miloš; Radotić, Ksenija

(Elsevier Science Bv, Amsterdam, 2016)

TY  - JOUR
AU  - Spasojević, Dragica
AU  - Zmejkoski, Danica
AU  - Glamočlija, Jasmina
AU  - Nikolić, Miloš
AU  - Soković, Marina
AU  - Milošević, Verica
AU  - Jarić, Ivana
AU  - Stojanović, Marijana M.
AU  - Marinković, Emilija
AU  - Barisani-Asenbauer, Talin
AU  - Prodanović, Radivoje
AU  - Jovanović, Miloš
AU  - Radotić, Ksenija
PY  - 2016
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/2364
AB  - Nowadays bacterial resistance to known antibiotics is a serious health problem. In order to achieve more efficient treatment, lately there is an effort to find new substances, such as certain biomaterials, that are non-toxic to humans with antibiotic potential. Lignins and lignin-derived compounds have been proposed to be good candidates for use in medicine and health maintenance. In this study, the antibacterial activity of the lignin model polymer dehydrogenate polymer (DHP) in alginate hydrogel (Alg) was studied. The obtained results show that DHP-Alg has strong antimicrobial activity against several bacterial strains and biofilms and does not have a toxic effect on human epithelial cells. These results strongly suggest its application as a wound healing agent or as an adjunct substance for wound treatments. (C) 2016 Elsevier B.V. and International Society of Chemotherapy. All rights reserved.
PB  - Elsevier Science Bv, Amsterdam
T2  - International Journal of Antimicrobial Agents
T1  - Lignin model compound in alginate hydrogel: a strong antimicrobial agent with high potential in wound treatment
VL  - 48
IS  - 6
SP  - 732
EP  - 735
DO  - 10.1016/j.ijantimicag.2016.08.014
ER  - 
@article{
author = "Spasojević, Dragica and Zmejkoski, Danica and Glamočlija, Jasmina and Nikolić, Miloš and Soković, Marina and Milošević, Verica and Jarić, Ivana and Stojanović, Marijana M. and Marinković, Emilija and Barisani-Asenbauer, Talin and Prodanović, Radivoje and Jovanović, Miloš and Radotić, Ksenija",
year = "2016",
abstract = "Nowadays bacterial resistance to known antibiotics is a serious health problem. In order to achieve more efficient treatment, lately there is an effort to find new substances, such as certain biomaterials, that are non-toxic to humans with antibiotic potential. Lignins and lignin-derived compounds have been proposed to be good candidates for use in medicine and health maintenance. In this study, the antibacterial activity of the lignin model polymer dehydrogenate polymer (DHP) in alginate hydrogel (Alg) was studied. The obtained results show that DHP-Alg has strong antimicrobial activity against several bacterial strains and biofilms and does not have a toxic effect on human epithelial cells. These results strongly suggest its application as a wound healing agent or as an adjunct substance for wound treatments. (C) 2016 Elsevier B.V. and International Society of Chemotherapy. All rights reserved.",
publisher = "Elsevier Science Bv, Amsterdam",
journal = "International Journal of Antimicrobial Agents",
title = "Lignin model compound in alginate hydrogel: a strong antimicrobial agent with high potential in wound treatment",
volume = "48",
number = "6",
pages = "732-735",
doi = "10.1016/j.ijantimicag.2016.08.014"
}
Spasojević, D., Zmejkoski, D., Glamočlija, J., Nikolić, M., Soković, M., Milošević, V., Jarić, I., Stojanović, M. M., Marinković, E., Barisani-Asenbauer, T., Prodanović, R., Jovanović, M.,& Radotić, K.. (2016). Lignin model compound in alginate hydrogel: a strong antimicrobial agent with high potential in wound treatment. in International Journal of Antimicrobial Agents
Elsevier Science Bv, Amsterdam., 48(6), 732-735.
https://doi.org/10.1016/j.ijantimicag.2016.08.014
Spasojević D, Zmejkoski D, Glamočlija J, Nikolić M, Soković M, Milošević V, Jarić I, Stojanović MM, Marinković E, Barisani-Asenbauer T, Prodanović R, Jovanović M, Radotić K. Lignin model compound in alginate hydrogel: a strong antimicrobial agent with high potential in wound treatment. in International Journal of Antimicrobial Agents. 2016;48(6):732-735.
doi:10.1016/j.ijantimicag.2016.08.014 .
Spasojević, Dragica, Zmejkoski, Danica, Glamočlija, Jasmina, Nikolić, Miloš, Soković, Marina, Milošević, Verica, Jarić, Ivana, Stojanović, Marijana M., Marinković, Emilija, Barisani-Asenbauer, Talin, Prodanović, Radivoje, Jovanović, Miloš, Radotić, Ksenija, "Lignin model compound in alginate hydrogel: a strong antimicrobial agent with high potential in wound treatment" in International Journal of Antimicrobial Agents, 48, no. 6 (2016):732-735,
https://doi.org/10.1016/j.ijantimicag.2016.08.014 . .
3
44
21
41
39

Supplementary data for the article: Spasojević, D.; Zmejkoski, D.; Glamočlija, J.; Nikolić, M.; Soković, M.; Milošević, V.; Jarić, I.; Stojanović, M.; Marinković, E.; Barisani-Asenbauer, T.; et al. Lignin Model Compound in Alginate Hydrogel: A Strong Antimicrobial Agent with High Potential in Wound Treatment. International Journal of Antimicrobial Agents 2016, 48 (6), 732–735. https://doi.org/10.1016/j.ijantimicag.2016.08.014

Spasojević, Dragica; Zmejkoski, Danica; Glamočlija, Jasmina; Nikolić, Miloš; Soković, Marina; Milošević, Verica; Jarić, Ivana; Stojanović, Marijana M.; Marinković, Emilija; Barisani-Asenbauer, Talin; Prodanović, Radivoje; Jovanović, Miloš; Radotić, Ksenija

(Elsevier Science Bv, Amsterdam, 2016)

TY  - DATA
AU  - Spasojević, Dragica
AU  - Zmejkoski, Danica
AU  - Glamočlija, Jasmina
AU  - Nikolić, Miloš
AU  - Soković, Marina
AU  - Milošević, Verica
AU  - Jarić, Ivana
AU  - Stojanović, Marijana M.
AU  - Marinković, Emilija
AU  - Barisani-Asenbauer, Talin
AU  - Prodanović, Radivoje
AU  - Jovanović, Miloš
AU  - Radotić, Ksenija
PY  - 2016
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/3560
PB  - Elsevier Science Bv, Amsterdam
T2  - International Journal of Antimicrobial Agents
T1  - Supplementary data for the article: Spasojević, D.; Zmejkoski, D.; Glamočlija, J.; Nikolić, M.; Soković, M.; Milošević, V.; Jarić, I.; Stojanović, M.; Marinković, E.; Barisani-Asenbauer, T.; et al. Lignin Model Compound in Alginate Hydrogel: A Strong Antimicrobial Agent with High Potential in Wound Treatment. International Journal of Antimicrobial Agents 2016, 48 (6), 732–735. https://doi.org/10.1016/j.ijantimicag.2016.08.014
UR  - https://hdl.handle.net/21.15107/rcub_cherry_3560
ER  - 
@misc{
author = "Spasojević, Dragica and Zmejkoski, Danica and Glamočlija, Jasmina and Nikolić, Miloš and Soković, Marina and Milošević, Verica and Jarić, Ivana and Stojanović, Marijana M. and Marinković, Emilija and Barisani-Asenbauer, Talin and Prodanović, Radivoje and Jovanović, Miloš and Radotić, Ksenija",
year = "2016",
publisher = "Elsevier Science Bv, Amsterdam",
journal = "International Journal of Antimicrobial Agents",
title = "Supplementary data for the article: Spasojević, D.; Zmejkoski, D.; Glamočlija, J.; Nikolić, M.; Soković, M.; Milošević, V.; Jarić, I.; Stojanović, M.; Marinković, E.; Barisani-Asenbauer, T.; et al. Lignin Model Compound in Alginate Hydrogel: A Strong Antimicrobial Agent with High Potential in Wound Treatment. International Journal of Antimicrobial Agents 2016, 48 (6), 732–735. https://doi.org/10.1016/j.ijantimicag.2016.08.014",
url = "https://hdl.handle.net/21.15107/rcub_cherry_3560"
}
Spasojević, D., Zmejkoski, D., Glamočlija, J., Nikolić, M., Soković, M., Milošević, V., Jarić, I., Stojanović, M. M., Marinković, E., Barisani-Asenbauer, T., Prodanović, R., Jovanović, M.,& Radotić, K.. (2016). Supplementary data for the article: Spasojević, D.; Zmejkoski, D.; Glamočlija, J.; Nikolić, M.; Soković, M.; Milošević, V.; Jarić, I.; Stojanović, M.; Marinković, E.; Barisani-Asenbauer, T.; et al. Lignin Model Compound in Alginate Hydrogel: A Strong Antimicrobial Agent with High Potential in Wound Treatment. International Journal of Antimicrobial Agents 2016, 48 (6), 732–735. https://doi.org/10.1016/j.ijantimicag.2016.08.014. in International Journal of Antimicrobial Agents
Elsevier Science Bv, Amsterdam..
https://hdl.handle.net/21.15107/rcub_cherry_3560
Spasojević D, Zmejkoski D, Glamočlija J, Nikolić M, Soković M, Milošević V, Jarić I, Stojanović MM, Marinković E, Barisani-Asenbauer T, Prodanović R, Jovanović M, Radotić K. Supplementary data for the article: Spasojević, D.; Zmejkoski, D.; Glamočlija, J.; Nikolić, M.; Soković, M.; Milošević, V.; Jarić, I.; Stojanović, M.; Marinković, E.; Barisani-Asenbauer, T.; et al. Lignin Model Compound in Alginate Hydrogel: A Strong Antimicrobial Agent with High Potential in Wound Treatment. International Journal of Antimicrobial Agents 2016, 48 (6), 732–735. https://doi.org/10.1016/j.ijantimicag.2016.08.014. in International Journal of Antimicrobial Agents. 2016;.
https://hdl.handle.net/21.15107/rcub_cherry_3560 .
Spasojević, Dragica, Zmejkoski, Danica, Glamočlija, Jasmina, Nikolić, Miloš, Soković, Marina, Milošević, Verica, Jarić, Ivana, Stojanović, Marijana M., Marinković, Emilija, Barisani-Asenbauer, Talin, Prodanović, Radivoje, Jovanović, Miloš, Radotić, Ksenija, "Supplementary data for the article: Spasojević, D.; Zmejkoski, D.; Glamočlija, J.; Nikolić, M.; Soković, M.; Milošević, V.; Jarić, I.; Stojanović, M.; Marinković, E.; Barisani-Asenbauer, T.; et al. Lignin Model Compound in Alginate Hydrogel: A Strong Antimicrobial Agent with High Potential in Wound Treatment. International Journal of Antimicrobial Agents 2016, 48 (6), 732–735. https://doi.org/10.1016/j.ijantimicag.2016.08.014" in International Journal of Antimicrobial Agents (2016),
https://hdl.handle.net/21.15107/rcub_cherry_3560 .

Recombinantly produced banana lectin isoform promotes balanced pro-inflammatory response in the colon

Marinković, Emilija; Lukić, Ivana; Kosanović, Dejana; Inić-Kanada, Aleksandra; Gavrović-Jankulović, Marija; Stojanović, Marijana M.

(Elsevier Science Bv, Amsterdam, 2016)

TY  - JOUR
AU  - Marinković, Emilija
AU  - Lukić, Ivana
AU  - Kosanović, Dejana
AU  - Inić-Kanada, Aleksandra
AU  - Gavrović-Jankulović, Marija
AU  - Stojanović, Marijana M.
PY  - 2016
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1934
AB  - Recombinant banana lectin isoform (rBanLec) attaches specifically to the mucosal surface, crosses the epithelial barrier and then directly affects the immune response in mouse colon. Structural characteristics, specificity and physiological impacts of rBanLec reported until now highly resemble those of its natural counterpart. Here, we demonstrated that a dose dependent stimulation of the colon with rBanLec skewed the immune response towards Th1/Th17 direction and this effect was counterbalanced by the rise in IL-10 production. Qualitative and quantitative characteristics of the established cytokine network were dependent on the applied rBanLec concentration. In addition, rBanLec enhanced local NO production and myeloperoxidase activity and promoted an increase in local IgA and IgG production. Stimulation with rBanLec can be beneficial in prevention of pathologies raised due to inappropriate cell-mediated immune response as well as in prevention of the pathogen invasion via the colon. (C) 2015 Elsevier Ltd. All rights reserved.
PB  - Elsevier Science Bv, Amsterdam
T2  - Journal of Functional Foods
T1  - Recombinantly produced banana lectin isoform promotes balanced pro-inflammatory response in the colon
VL  - 20
SP  - 68
EP  - 78
DO  - 10.1016/j.jff.2015.10.019
ER  - 
@article{
author = "Marinković, Emilija and Lukić, Ivana and Kosanović, Dejana and Inić-Kanada, Aleksandra and Gavrović-Jankulović, Marija and Stojanović, Marijana M.",
year = "2016",
abstract = "Recombinant banana lectin isoform (rBanLec) attaches specifically to the mucosal surface, crosses the epithelial barrier and then directly affects the immune response in mouse colon. Structural characteristics, specificity and physiological impacts of rBanLec reported until now highly resemble those of its natural counterpart. Here, we demonstrated that a dose dependent stimulation of the colon with rBanLec skewed the immune response towards Th1/Th17 direction and this effect was counterbalanced by the rise in IL-10 production. Qualitative and quantitative characteristics of the established cytokine network were dependent on the applied rBanLec concentration. In addition, rBanLec enhanced local NO production and myeloperoxidase activity and promoted an increase in local IgA and IgG production. Stimulation with rBanLec can be beneficial in prevention of pathologies raised due to inappropriate cell-mediated immune response as well as in prevention of the pathogen invasion via the colon. (C) 2015 Elsevier Ltd. All rights reserved.",
publisher = "Elsevier Science Bv, Amsterdam",
journal = "Journal of Functional Foods",
title = "Recombinantly produced banana lectin isoform promotes balanced pro-inflammatory response in the colon",
volume = "20",
pages = "68-78",
doi = "10.1016/j.jff.2015.10.019"
}
Marinković, E., Lukić, I., Kosanović, D., Inić-Kanada, A., Gavrović-Jankulović, M.,& Stojanović, M. M.. (2016). Recombinantly produced banana lectin isoform promotes balanced pro-inflammatory response in the colon. in Journal of Functional Foods
Elsevier Science Bv, Amsterdam., 20, 68-78.
https://doi.org/10.1016/j.jff.2015.10.019
Marinković E, Lukić I, Kosanović D, Inić-Kanada A, Gavrović-Jankulović M, Stojanović MM. Recombinantly produced banana lectin isoform promotes balanced pro-inflammatory response in the colon. in Journal of Functional Foods. 2016;20:68-78.
doi:10.1016/j.jff.2015.10.019 .
Marinković, Emilija, Lukić, Ivana, Kosanović, Dejana, Inić-Kanada, Aleksandra, Gavrović-Jankulović, Marija, Stojanović, Marijana M., "Recombinantly produced banana lectin isoform promotes balanced pro-inflammatory response in the colon" in Journal of Functional Foods, 20 (2016):68-78,
https://doi.org/10.1016/j.jff.2015.10.019 . .
4
5
6
4

Recombinant banana lectin as mucosal immunostimulator

Dimitrijevic, R.; Stojanovic, M.; Micic, M.; Dimitrijevic, Lj; Gavrović-Jankulović, Marija

(Elsevier Science Bv, Amsterdam, 2012)

TY  - JOUR
AU  - Dimitrijevic, R.
AU  - Stojanovic, M.
AU  - Micic, M.
AU  - Dimitrijevic, Lj
AU  - Gavrović-Jankulović, Marija
PY  - 2012
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1310
AB  - To explore the potential usage of recombinant banana lectin as an immunostimulator it was of interest to analyze the effect it exerted in vivo in mice, upon two different routes of exposure. By Western blotting analysis of the gastric and intestinal luminal content it was found that recombinant banana lectin is stable in vivo in the mouse digestive tract, where it specifically interacts with the mucosal surfaces which can be inhibited by the addition of glucose. Its attachment to mice jejunum has been confirmed by immunofluorescence. In addition, it was able to induce systemic immunity, evidenced by specific antibody production, by oral application, a property that might be exploited for the induction of systemic immune responses. (c) 2012 Elsevier Ltd. All rights reserved.
PB  - Elsevier Science Bv, Amsterdam
T2  - Journal of Functional Foods
T1  - Recombinant banana lectin as mucosal immunostimulator
VL  - 4
IS  - 3
SP  - 636
EP  - 641
DO  - 10.1016/j.jff.2012.04.003
ER  - 
@article{
author = "Dimitrijevic, R. and Stojanovic, M. and Micic, M. and Dimitrijevic, Lj and Gavrović-Jankulović, Marija",
year = "2012",
abstract = "To explore the potential usage of recombinant banana lectin as an immunostimulator it was of interest to analyze the effect it exerted in vivo in mice, upon two different routes of exposure. By Western blotting analysis of the gastric and intestinal luminal content it was found that recombinant banana lectin is stable in vivo in the mouse digestive tract, where it specifically interacts with the mucosal surfaces which can be inhibited by the addition of glucose. Its attachment to mice jejunum has been confirmed by immunofluorescence. In addition, it was able to induce systemic immunity, evidenced by specific antibody production, by oral application, a property that might be exploited for the induction of systemic immune responses. (c) 2012 Elsevier Ltd. All rights reserved.",
publisher = "Elsevier Science Bv, Amsterdam",
journal = "Journal of Functional Foods",
title = "Recombinant banana lectin as mucosal immunostimulator",
volume = "4",
number = "3",
pages = "636-641",
doi = "10.1016/j.jff.2012.04.003"
}
Dimitrijevic, R., Stojanovic, M., Micic, M., Dimitrijevic, L.,& Gavrović-Jankulović, M.. (2012). Recombinant banana lectin as mucosal immunostimulator. in Journal of Functional Foods
Elsevier Science Bv, Amsterdam., 4(3), 636-641.
https://doi.org/10.1016/j.jff.2012.04.003
Dimitrijevic R, Stojanovic M, Micic M, Dimitrijevic L, Gavrović-Jankulović M. Recombinant banana lectin as mucosal immunostimulator. in Journal of Functional Foods. 2012;4(3):636-641.
doi:10.1016/j.jff.2012.04.003 .
Dimitrijevic, R., Stojanovic, M., Micic, M., Dimitrijevic, Lj, Gavrović-Jankulović, Marija, "Recombinant banana lectin as mucosal immunostimulator" in Journal of Functional Foods, 4, no. 3 (2012):636-641,
https://doi.org/10.1016/j.jff.2012.04.003 . .
11
11
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10

In vitro stimulation of Balb/c and C57 BL/6 splenocytes by a recombinantly produced banana lectin isoform results in both a proliferation of T cells and an increased secretion of interferon-gamma

Stojanović, Marijana M.; Živković, Irena; Petrusic, Vladimir Z.; Kosec, Dusko J.; Dimitrijevic, Rajna D.; Jankov, Ratko M.; Dimitrijevic, Ljijana A.; Gavrović-Jankulović, Marija

(Elsevier Science Bv, Amsterdam, 2010)

TY  - JOUR
AU  - Stojanović, Marijana M.
AU  - Živković, Irena
AU  - Petrusic, Vladimir Z.
AU  - Kosec, Dusko J.
AU  - Dimitrijevic, Rajna D.
AU  - Jankov, Ratko M.
AU  - Dimitrijevic, Ljijana A.
AU  - Gavrović-Jankulović, Marija
PY  - 2010
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1043
AB  - Lectins are widely used in many types of assay but some lectins such as banana lectin (BanLec) are recognised as potent immunostimulators Although BanLec's structure and binding characteristics are now familiar, its immunostimulatory potential has not yet been fully explored The synthesis by recombinant technology of a BanLec isoform (rBanLec) whose binding properties are similar to its natural counterpart has made it possible to overcome the twin problems of natural BanLec's microheterogeneity and low availability This study's aim is to explore the immunostimulatory potential of rBanLec in the murine model Analyses of the responses of Balb/c- and C57 BL/6-originated splenocytes to in vitro rBanLec stimulation were performed to examine the dependency of rBanLec's immunostimulatory potential upon the splenocytes' genetic background It is shown that the responses of Balb/c- and C57 BL/6-originated splenocytes to rBanLec stimulation differ both qualitatively and in intensity. The hallmarks of the induced responses are T lymphocyte proliferation and intensive interferon-gamma secretion Both phenomena are more marked in Balb/c-originated cultures; Balb/c-originated lymphocytes produce interleukin (IL)-4 and IL-10 following rBanLec stimulation Out results demonstrate that any responses to rBanLec stimulation are highly dependent upon genetic background. they suggest that genetic background must be an important consideration in any further investigations using animal models or when exploring rBanLec's potential human applications (C) 2009 Elsevier B V. All rights reserved
PB  - Elsevier Science Bv, Amsterdam
T2  - International Immunopharmacology
T1  - In vitro stimulation of Balb/c and C57 BL/6 splenocytes by a recombinantly produced banana lectin isoform results in both a proliferation of T cells and an increased secretion of interferon-gamma
VL  - 10
IS  - 1
SP  - 120
EP  - 129
DO  - 10.1016/j.intimp.2009.10.007
ER  - 
@article{
author = "Stojanović, Marijana M. and Živković, Irena and Petrusic, Vladimir Z. and Kosec, Dusko J. and Dimitrijevic, Rajna D. and Jankov, Ratko M. and Dimitrijevic, Ljijana A. and Gavrović-Jankulović, Marija",
year = "2010",
abstract = "Lectins are widely used in many types of assay but some lectins such as banana lectin (BanLec) are recognised as potent immunostimulators Although BanLec's structure and binding characteristics are now familiar, its immunostimulatory potential has not yet been fully explored The synthesis by recombinant technology of a BanLec isoform (rBanLec) whose binding properties are similar to its natural counterpart has made it possible to overcome the twin problems of natural BanLec's microheterogeneity and low availability This study's aim is to explore the immunostimulatory potential of rBanLec in the murine model Analyses of the responses of Balb/c- and C57 BL/6-originated splenocytes to in vitro rBanLec stimulation were performed to examine the dependency of rBanLec's immunostimulatory potential upon the splenocytes' genetic background It is shown that the responses of Balb/c- and C57 BL/6-originated splenocytes to rBanLec stimulation differ both qualitatively and in intensity. The hallmarks of the induced responses are T lymphocyte proliferation and intensive interferon-gamma secretion Both phenomena are more marked in Balb/c-originated cultures; Balb/c-originated lymphocytes produce interleukin (IL)-4 and IL-10 following rBanLec stimulation Out results demonstrate that any responses to rBanLec stimulation are highly dependent upon genetic background. they suggest that genetic background must be an important consideration in any further investigations using animal models or when exploring rBanLec's potential human applications (C) 2009 Elsevier B V. All rights reserved",
publisher = "Elsevier Science Bv, Amsterdam",
journal = "International Immunopharmacology",
title = "In vitro stimulation of Balb/c and C57 BL/6 splenocytes by a recombinantly produced banana lectin isoform results in both a proliferation of T cells and an increased secretion of interferon-gamma",
volume = "10",
number = "1",
pages = "120-129",
doi = "10.1016/j.intimp.2009.10.007"
}
Stojanović, M. M., Živković, I., Petrusic, V. Z., Kosec, D. J., Dimitrijevic, R. D., Jankov, R. M., Dimitrijevic, L. A.,& Gavrović-Jankulović, M.. (2010). In vitro stimulation of Balb/c and C57 BL/6 splenocytes by a recombinantly produced banana lectin isoform results in both a proliferation of T cells and an increased secretion of interferon-gamma. in International Immunopharmacology
Elsevier Science Bv, Amsterdam., 10(1), 120-129.
https://doi.org/10.1016/j.intimp.2009.10.007
Stojanović MM, Živković I, Petrusic VZ, Kosec DJ, Dimitrijevic RD, Jankov RM, Dimitrijevic LA, Gavrović-Jankulović M. In vitro stimulation of Balb/c and C57 BL/6 splenocytes by a recombinantly produced banana lectin isoform results in both a proliferation of T cells and an increased secretion of interferon-gamma. in International Immunopharmacology. 2010;10(1):120-129.
doi:10.1016/j.intimp.2009.10.007 .
Stojanović, Marijana M., Živković, Irena, Petrusic, Vladimir Z., Kosec, Dusko J., Dimitrijevic, Rajna D., Jankov, Ratko M., Dimitrijevic, Ljijana A., Gavrović-Jankulović, Marija, "In vitro stimulation of Balb/c and C57 BL/6 splenocytes by a recombinantly produced banana lectin isoform results in both a proliferation of T cells and an increased secretion of interferon-gamma" in International Immunopharmacology, 10, no. 1 (2010):120-129,
https://doi.org/10.1016/j.intimp.2009.10.007 . .
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The identification of a low molecular mass bacteriocin, rhamnosin A, produced by Lactobacillus rhamnosus strain 68

Dimitrijevic, R.; Stojanovic, M.; Živković, Irena; Petersen, A.; Jankov, Ratko M.; Dimitrijevic, L.; Gavrović-Jankulović, Marija

(Wiley-Blackwell, Hoboken, 2009)

TY  - JOUR
AU  - Dimitrijevic, R.
AU  - Stojanovic, M.
AU  - Živković, Irena
AU  - Petersen, A.
AU  - Jankov, Ratko M.
AU  - Dimitrijevic, L.
AU  - Gavrović-Jankulović, Marija
PY  - 2009
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1028
AB  - Aims: This study focuses on the isolation and characterization of a peptide with bacteriocin-like properties isolated from Lactobacillus rhamnosus strain 68, previously identified by 16S rRNA gene sequencing and originating from human gastrointestinal flora. Methods and Results: The peptide was isolated from a supernatant of bacteria maintained under restrictive conditions by a combination of ethanol precipitation and reversed-phase chromatography. The molecular mass of the peptide as assessed by mass spectrometry was 6433 center dot 8 Da. An isoelectric point of 9 center dot 8 was determined by 2D-PAGE. The peptide designated rhamnosin A inhibited Micrococcus lysodeikticus ATCC 4698 but did not inhibit Lactobacillus plantarum 8014 or Lact. plantarum 39268. Inhibitory activity against M. lysodeikticus at concentrations used in this study was shown to be bacteriostatic rather than bacteriolytic or bactericidal. Rhamnosin A retained biological activity after heat treatment (95 degrees C, 30 min) but was sensitive to proteolytic activity of pepsin and trypsin. Conclusions: The N-terminal sequence of rhamnosin A, as determined by Edman degradation and in more detail by blast analysis, did not show identity with any currently available Lact. rhamnosus HN001-translated protein sequences, nor any significant similarity with other sequences in the nonredundant protein sequence database. Being a small, heat-stable, nonlanthionine-containing peptide, rhamnosin A should be categorized as a class II bacteriocin. Significance and Impact of the Study: This study describes a partial bacteriocin sequence isolated from Lact. rhamnosus 68 and broadens our understanding of bacteriocins.
PB  - Wiley-Blackwell, Hoboken
T2  - Journal of Applied Microbiology
T1  - The identification of a low molecular mass bacteriocin, rhamnosin A, produced by Lactobacillus rhamnosus strain 68
VL  - 107
IS  - 6
SP  - 2108
EP  - 2115
DO  - 10.1111/j.1365-2672.2009.04539.x
ER  - 
@article{
author = "Dimitrijevic, R. and Stojanovic, M. and Živković, Irena and Petersen, A. and Jankov, Ratko M. and Dimitrijevic, L. and Gavrović-Jankulović, Marija",
year = "2009",
abstract = "Aims: This study focuses on the isolation and characterization of a peptide with bacteriocin-like properties isolated from Lactobacillus rhamnosus strain 68, previously identified by 16S rRNA gene sequencing and originating from human gastrointestinal flora. Methods and Results: The peptide was isolated from a supernatant of bacteria maintained under restrictive conditions by a combination of ethanol precipitation and reversed-phase chromatography. The molecular mass of the peptide as assessed by mass spectrometry was 6433 center dot 8 Da. An isoelectric point of 9 center dot 8 was determined by 2D-PAGE. The peptide designated rhamnosin A inhibited Micrococcus lysodeikticus ATCC 4698 but did not inhibit Lactobacillus plantarum 8014 or Lact. plantarum 39268. Inhibitory activity against M. lysodeikticus at concentrations used in this study was shown to be bacteriostatic rather than bacteriolytic or bactericidal. Rhamnosin A retained biological activity after heat treatment (95 degrees C, 30 min) but was sensitive to proteolytic activity of pepsin and trypsin. Conclusions: The N-terminal sequence of rhamnosin A, as determined by Edman degradation and in more detail by blast analysis, did not show identity with any currently available Lact. rhamnosus HN001-translated protein sequences, nor any significant similarity with other sequences in the nonredundant protein sequence database. Being a small, heat-stable, nonlanthionine-containing peptide, rhamnosin A should be categorized as a class II bacteriocin. Significance and Impact of the Study: This study describes a partial bacteriocin sequence isolated from Lact. rhamnosus 68 and broadens our understanding of bacteriocins.",
publisher = "Wiley-Blackwell, Hoboken",
journal = "Journal of Applied Microbiology",
title = "The identification of a low molecular mass bacteriocin, rhamnosin A, produced by Lactobacillus rhamnosus strain 68",
volume = "107",
number = "6",
pages = "2108-2115",
doi = "10.1111/j.1365-2672.2009.04539.x"
}
Dimitrijevic, R., Stojanovic, M., Živković, I., Petersen, A., Jankov, R. M., Dimitrijevic, L.,& Gavrović-Jankulović, M.. (2009). The identification of a low molecular mass bacteriocin, rhamnosin A, produced by Lactobacillus rhamnosus strain 68. in Journal of Applied Microbiology
Wiley-Blackwell, Hoboken., 107(6), 2108-2115.
https://doi.org/10.1111/j.1365-2672.2009.04539.x
Dimitrijevic R, Stojanovic M, Živković I, Petersen A, Jankov RM, Dimitrijevic L, Gavrović-Jankulović M. The identification of a low molecular mass bacteriocin, rhamnosin A, produced by Lactobacillus rhamnosus strain 68. in Journal of Applied Microbiology. 2009;107(6):2108-2115.
doi:10.1111/j.1365-2672.2009.04539.x .
Dimitrijevic, R., Stojanovic, M., Živković, Irena, Petersen, A., Jankov, Ratko M., Dimitrijevic, L., Gavrović-Jankulović, Marija, "The identification of a low molecular mass bacteriocin, rhamnosin A, produced by Lactobacillus rhamnosus strain 68" in Journal of Applied Microbiology, 107, no. 6 (2009):2108-2115,
https://doi.org/10.1111/j.1365-2672.2009.04539.x . .
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Quantification of the thaumatin-like kiwi allergen by a monoclonal antibody-based ELISA

Gavrović-Jankulović, Marija; Spasic, Milena; Ćirković-Veličković, Tanja; Stojanović, Marijana M.; Inić-Kanada, Aleksandra; Dimitrijevic, Ljiliana; Lindner, Buko; Petersen, Arnd; Becker, Wolf-Meinhard; Jankov, Ratko M.

(Wiley-Blackwell, Malden, 2008)

TY  - JOUR
AU  - Gavrović-Jankulović, Marija
AU  - Spasic, Milena
AU  - Ćirković-Veličković, Tanja
AU  - Stojanović, Marijana M.
AU  - Inić-Kanada, Aleksandra
AU  - Dimitrijevic, Ljiliana
AU  - Lindner, Buko
AU  - Petersen, Arnd
AU  - Becker, Wolf-Meinhard
AU  - Jankov, Ratko M.
PY  - 2008
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/948
AB  - Thaumatin-like proteins (TLPs) have been established as a new family of fruit and pollen allergens. The aim of this study was to develop a two-site ELISA for the quantification of the thaumatin-like kiwi allergen (Act d 2) in kiwifruit extracts and kiwi fruit-containing food products. Genomic DNA (gDNA) of Act d 2 was amplified and the deduced amino acid sequence was determined to obtain a primary structure. Act d 2 purified from kiwifruit extract by HPLC was identified by Edman degradation and MS. Balb/c mice were immunized with Act d 2 for the production of mAbs by hybridoma technology. The optimized ELISA measured Act d 2 concentrations ranging from 0.2 to 9.0 ng/mL, with intra- and interassay coefficients of variation of 3.65 and 10.44%, respectively. The developed ELISA is a useful method for the quantification of the thaumatin-like kiwi allergen in kiwifruit extracts as well as the allergen level in kiwifruit-containing food products. It may be a helpful analytical tool for the evaluation of the stability (integrity) of fruit allergen extracts for in vitro diagnosis.
PB  - Wiley-Blackwell, Malden
T2  - Molecular Nutrition and Food Research
T1  - Quantification of the thaumatin-like kiwi allergen by a monoclonal antibody-based ELISA
VL  - 52
IS  - 6
SP  - 701
EP  - 707
DO  - 10.1002/mnfr.200700286
ER  - 
@article{
author = "Gavrović-Jankulović, Marija and Spasic, Milena and Ćirković-Veličković, Tanja and Stojanović, Marijana M. and Inić-Kanada, Aleksandra and Dimitrijevic, Ljiliana and Lindner, Buko and Petersen, Arnd and Becker, Wolf-Meinhard and Jankov, Ratko M.",
year = "2008",
abstract = "Thaumatin-like proteins (TLPs) have been established as a new family of fruit and pollen allergens. The aim of this study was to develop a two-site ELISA for the quantification of the thaumatin-like kiwi allergen (Act d 2) in kiwifruit extracts and kiwi fruit-containing food products. Genomic DNA (gDNA) of Act d 2 was amplified and the deduced amino acid sequence was determined to obtain a primary structure. Act d 2 purified from kiwifruit extract by HPLC was identified by Edman degradation and MS. Balb/c mice were immunized with Act d 2 for the production of mAbs by hybridoma technology. The optimized ELISA measured Act d 2 concentrations ranging from 0.2 to 9.0 ng/mL, with intra- and interassay coefficients of variation of 3.65 and 10.44%, respectively. The developed ELISA is a useful method for the quantification of the thaumatin-like kiwi allergen in kiwifruit extracts as well as the allergen level in kiwifruit-containing food products. It may be a helpful analytical tool for the evaluation of the stability (integrity) of fruit allergen extracts for in vitro diagnosis.",
publisher = "Wiley-Blackwell, Malden",
journal = "Molecular Nutrition and Food Research",
title = "Quantification of the thaumatin-like kiwi allergen by a monoclonal antibody-based ELISA",
volume = "52",
number = "6",
pages = "701-707",
doi = "10.1002/mnfr.200700286"
}
Gavrović-Jankulović, M., Spasic, M., Ćirković-Veličković, T., Stojanović, M. M., Inić-Kanada, A., Dimitrijevic, L., Lindner, B., Petersen, A., Becker, W.,& Jankov, R. M.. (2008). Quantification of the thaumatin-like kiwi allergen by a monoclonal antibody-based ELISA. in Molecular Nutrition and Food Research
Wiley-Blackwell, Malden., 52(6), 701-707.
https://doi.org/10.1002/mnfr.200700286
Gavrović-Jankulović M, Spasic M, Ćirković-Veličković T, Stojanović MM, Inić-Kanada A, Dimitrijevic L, Lindner B, Petersen A, Becker W, Jankov RM. Quantification of the thaumatin-like kiwi allergen by a monoclonal antibody-based ELISA. in Molecular Nutrition and Food Research. 2008;52(6):701-707.
doi:10.1002/mnfr.200700286 .
Gavrović-Jankulović, Marija, Spasic, Milena, Ćirković-Veličković, Tanja, Stojanović, Marijana M., Inić-Kanada, Aleksandra, Dimitrijevic, Ljiliana, Lindner, Buko, Petersen, Arnd, Becker, Wolf-Meinhard, Jankov, Ratko M., "Quantification of the thaumatin-like kiwi allergen by a monoclonal antibody-based ELISA" in Molecular Nutrition and Food Research, 52, no. 6 (2008):701-707,
https://doi.org/10.1002/mnfr.200700286 . .
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