Abstract
As severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) continues to spread globally, a series of vaccines, antibodies and drugs have been developed to combat coronavirus disease 2019 (COVID-19). High specific antibodies are powerful tool for the development of immunoassay and providing passive immunotherapy against SARS-CoV-2 and expected with large scale production. SARS-CoV-2 S1 protein was expressed in E. coli BL21 and purified by immobilized metal affinity chromatography, as antigen used to immunize hens, the specific IgY antibodies were extracted form egg yolk by PEG-6000 precipitation, and the titer of anti-S1 IgY antibody reached 1:10,000. IgY single chain variable fragment antibody (IgY-scFv) was generated by using phage display technology and the IgY-scFv showed high binding sensitivity and capacity to S1 protein of SARS-CoV-2, and the minimum detectable antigen S1 protein concentration was 6 ng/µL. The docking study showed that the multiple epitopes on the IgY-scFv interacted with multiple residues on SARS-CoV-2 S1 RBD to form hydrogen bonds. This preliminary study suggests that IgY and IgY-scFv are suitable candidates for the development of immunoassay and passive immunotherapy for COVID-19 to humans and animals.
Highlights
COVID-19, first reported in December 2019, can cause pneumonia-like symptoms, including fever, cough and fatigue
Antibodies are the powerful tool for the development of point-of-care testing (POCT) immunoassay in SARSCoV-2 analysis, the desired method should be accurate, specific, rapid, robust and inexpensive (Seo et al 2020; Kim et al 2021)
This study aimed to generate and evaluate the IgYscFv against SARS-CoV-2 S1 protein, which may offer an alternative solution in providing large number of highlyspecific antibodies in combating COVID-19 in different application scenarios
Summary
COVID-19, first reported in December 2019, can cause pneumonia-like symptoms, including fever, cough and fatigue. The pathogen, SARS-CoV-2, is positive-sense singlestranded RNA virus with a genome of approximately 30 kbp encodes four structural proteins, including spike, envelop, matrix and nucleocapsid (Kim et al 2021). The spike (S) protein comprises two components: S1, which contains the distinct receptor binding domain (RBD); S2, which contains the fusion peptide. Antibodies against the S1 subunit, targeting to RBD, are expected to prevent the viral particle surface protein from binding to the ACE2 receptor, preventing viral invasion (Parray et al 2020). The SARS-CoV-2 S1 protein has been accepted as ideal target for antibody and vaccine design (Premkumar et al 2020; Parray et al 2020). Antibodies are the powerful tool for the development of point-of-care testing (POCT) immunoassay in SARSCoV-2 analysis, the desired method should be accurate, specific, rapid, robust and inexpensive (Seo et al 2020; Kim et al 2021)
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