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Proteomic and immunological characterization of recombinantly expressed nucleocapsid SARS-CoV 2 protein fragment in E. coli
dc.creator | Đukić, Teodora | |
dc.creator | Mladenović, Maja | |
dc.creator | Stanić-Vučinić, Dragana | |
dc.creator | Radosavljević, Jelena | |
dc.creator | Smiljanić, Katarina | |
dc.creator | Sabljić, Ljiljana | |
dc.creator | Gnjatović, Marija Lj. | |
dc.creator | Cujić, Danica | |
dc.creator | Vasović, Tamara | |
dc.creator | Simović, Ana | |
dc.creator | Radomirović, Mirjana Ž. | |
dc.creator | Ćirković-Veličković, Tanja | |
dc.date.accessioned | 2022-12-29T17:40:44Z | |
dc.date.available | 2022-12-29T17:40:44Z | |
dc.date.issued | 2021 | |
dc.identifier.uri | http://cherry.chem.bg.ac.rs/handle/123456789/5735 | |
dc.description.abstract | Serological testing is important method for diagnosis of severe acute respiratory syndrome coronavirus 2 (SARSCoV-2) infection and for assessment of immunological response after the vaccination. Nucleocapsid (N) protein is the most abundant virus protein and strong immunogen. The aim was develop efficient, low-cost production of N protein large fragment and to characterize it with bottom-up, high-resolution tandem mass spectrometry and immunologically. SARS-CoV-2 recombinant fragment of nucleocapsid protein (rfNP; 58–419 aa) was expressed in E. coli in soluble form and purified by several chromatographic steps and was subjected to SDS-PAGE and in-gel digested with trypsin. rfNP was tested in immunoblot using sera of COVID-19 convalescent patients. ELISA was optimized with sera of RT-PCR confirmed positive symptomatic patients and healthy individuals. IgG detection sensitivity was 96% (47/50) and specificity 97% (67/68), while IgM detection was slightly lower. Identity of rfNP was confirmed with high scores and peptide coverage above 80%. Estimation from the value of areas under ion extracted chromatographic curves is that only up to 0,03% of the total band protein quantity belongs to host proteins, while rfNP share is well above 99,9%, resulting in highly pure nucleocapsid protein preparation. | sr |
dc.language.iso | en | sr |
dc.relation | info:eu-repo/grantAgreement/MESTD/inst-2020/200168/RS// | sr |
dc.relation | info:eu-repo/grantAgreement/EC/H2020/810752/EU// | sr |
dc.relation | info:eu-repo/grantAgreement/ScienceFundRS/Fond_2020_COVID19/7542203/RS// | sr |
dc.relation | 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. | sr |
dc.rights | openAccess | sr |
dc.rights.uri | https://creativecommons.org/licenses/by/4.0/ | |
dc.source | Proteomics and Metabolomics for Personalized Medicine, XV Italian Proteomics Association Annual Meeting, Catholic University of the Sacred Heart, Roma, Italy, 8th-10th September 2021 | sr |
dc.subject | COVID-19 | sr |
dc.subject | recombinant protein | sr |
dc.subject | nucleocapsid | sr |
dc.subject | Prokaryotic expression | sr |
dc.subject | SARS-CoV-2 | sr |
dc.subject | serological assay | sr |
dc.title | Proteomic and immunological characterization of recombinantly expressed nucleocapsid SARS-CoV 2 protein fragment in E. coli | sr |
dc.type | conferenceObject | sr |
dc.rights.license | BY | sr |
dc.citation.spage | 50 | |
dc.citation.epage | 50 | |
dc.description.other | Book of Abstracts | sr |
dc.type.version | publishedVersion | sr |
dc.identifier.fulltext | http://cherry.chem.bg.ac.rs/bitstream/id/32201/bitstream_32201.pdf | |
dc.identifier.rcub | https://hdl.handle.net/21.15107/rcub_cherry_5735 |