Development and ELISA-based detection of anti-M2e IgY antibodies using an encoding plasmid for M2e-Hsp70 C-terminal gene

Abstract

Background: The use of IgYs in a variety of methods in different areas of research, diagnostics, medical application and biotechnology should be considered widely. Objectives: Development of antibodies against extra cellular domain of influenza M2 (M2e) protein in egg yolk of laying hens. Methods: A Fusion construct harboring C-terminal of bovine heat shock protein 70 (Hsp70) and influenza M2e coding genes was injected to laying hens. Serum and egg yolk antibodies were screened for the presence of anti-M2e antibodies by indirect enzyme-linked immunosorbent assay (ELISA). Results: Anti-M2e antibodies were detected in egg yolks and sera of injected hens from 13 and 7 days post injection (PI), with the peak titer detected on 41 and 35 days PI, respectively. Conclusions: Anti-M2e IgY titers could be an index for expression potential of pcDNA3.1-M2e-HspC-terminal construct in laying hens. This construct could be considered as a promising tool in production of anti-M2e polyclonal, monospecific IgY antibodies. Such anti-M2e antibodies could be exploited for influenza diagnostic and therapeutic measures.