Proteomic identification of recombinant nucleocapsid N protein 40 kDa fragment from SARS-CoV-2

ABSTRACT: Serological testing is important methodfor diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Nucleocapsid (N) protein is the most abundant virus derived protein and strong immunogen. We aimed to find its efficient, low-cost production. SARS-CoV-2 recombinant fragment of nucleocapsid protein (rfNP; 58-419 aa) was expressed in E. coli in soluble form, purified and characterized biochemically and immunologically. Purified rfNP has secondary structure of full-length recombinant N protein, with high percentage of disordered structure (34.2 %) and of β-sheet (40.7 %). rfNP was tested in immunoblot using sera of COVID-19 convalescent patients. ELISA was optimized with sera of RT-PCR confirmed positive symptomatic patients and healthy individuals. IgG detection sensitivity was 96% (47/50) and specificity 97% (67/68), while IgM detection was slightly lower (94% and 96.5%, respectively). Cost-effective approach for soluble recombinant N protein fragment production was developed, with reliable IgG and IgM antibodies detection of SARS-CoV-2 infection.

INSTRUMENT(S): LTQ Orbitrap XL

ORGANISM(S): Sars Coronavirus Sin3765v

TISSUE(S): Organism Or Virus Or Viroid

DISEASE(S): Covid-19

SUBMITTER: Katarina Smiljanic  

LAB HEAD: Tanja D. Ćirković Velicković

PROVIDER: PXD023341 | Pride | 2021-02-17

REPOSITORIES: Pride