Abstract
Coronavirus disease (COVID)-19 caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection has become a global pandemic disease that has social and economic chaos. An alternative mitigation strategy may involve the use of specific immunoglobulin (Ig)-Y derived from chicken eggs. Our study aimed to evaluate the neutralizing potential of specific IgY targeting S1, receptor-binding-domain (RBD) of spike glycoprotein and nucleocapsid (N) of SARS-CoV-2 to inhibit RBD and angiotensin-converting-enzyme-2 (ACE2) binding interaction. Hy-Line Brown laying hens were immunized with recombinant S1, RBD spike glycoprotein, and nucleocapsid (N) of SARS-CoV-2. The presence of specific S1,RBD,N-IgY in serum and egg yolk was verified by indirect enzyme-linked immunosorbent assay (ELISA). Specific S1,RBD,N-IgY was purified and characterized from egg yolk using sodium-dodecyl-sulfate-polyacrylamide-gel-electrophoresis (SDS-PAGE), and was subsequently evaluated for inhibition of the RBD-ACE2 binding interaction in vitro. Specific IgY was present in serum at 1 week post-initial immunization (p.i.i), whereas its present in egg yolk was confirmed at 4 weeks p.i.i. Specific S1,RBD,N-IgY in serum was able to inhibit RBD-ACE2 binding interaction between 4 and 15 weeks p.i.i. The results of the SDS-PAGE revealed the presence of bands with molecular weights of 180 kDa, indicating the presence of whole IgY. Our results demonstrated that S1,RBD,N-IgY was able to inhibit RBD-ACE2 binding interaction in vitro, suggesting its potential use in blocking virus entry. Our study also demonstrated proof-of-concept that laying hens were able to produce this specific IgY, which could block the viral binding and large production of this specific IgY is feasible.
Keywords: Angiotensin-converting-enzyme-2; Binding interaction; Immunoglobulin Y; Laying hens; Receptor-binding-domain; SARS-CoV-2.
【저자키워드】 SARS-CoV-2., immunoglobulin Y, Receptor-binding-domain, Angiotensin-converting-enzyme-2, binding interaction, Laying hens, 【초록키워드】 SARS-CoV-2, ACE2, coronavirus, spike glycoprotein, Infection, in vitro, immunization, ELISA, global pandemic, serum, Immunoglobulin, nucleocapsid, RBD, Neutralizing, disease, binding, acute respiratory syndrome, Specific, Brown, molecular weight, immunized, enzyme-linked immunosorbent, SDS-PAGE, blocking virus entry, RBD-ACE2, evaluate, caused, inhibit, evaluated, characterized, demonstrated, kDa, feasible, purified, Laying, 【제목키워드】 SARS-CoV-2, spike glycoprotein, Receptor binding domain, nucleocapsid, RBD-ACE2,