Exploration on a new routine for preparing rabies laboratory diagnostic monoclonal antibodies

Abstract

Objective To explore the possibility of preparing rabies laboratory diagnostic monoclonal antibodies with synthetic peptides as immunogen.Methods A synthetic peptide located at residues 355 to 369 of rabies virus CVS-11 nucleoprotein was conjugated to the Keyhole Limpe hemocyanin (KLH) macromolecule and used to immunize BALB/c mice to prepare hybridoma cell lines by classical hybridoma technology.Indirect enzyme-linked immunosorbent assay (ELISA) and indirect fluorescent assay (IFA) were used to screen and identify the hybridoma cell lines.Results A positive hybridoma cell line named 2B1D11 was obtained after screening the anti-peptide antibodies in supernatants of hybridoma cell lines by indirect ELISA and IFA.Monoclonal antibody in ascites of BALB/c mice inoculated with 2B1D11 was purified and used successfully to detect the rabies virus in canine brain tissues and BHK-21 cells by IFA.Conclusion It is possible to prepare rabies laboratory diagnostic monoclonal antibodies with synthetic peptides as immunogen. Key words: Rabies virus; Nucleoproteins ; Peptides; Antibody/diagnosis