ABSTRACT
Single radial immunodiffusion (SRD) assays were used for measuring the haemagglutinin antigen contents of equine influenza vaccine prepared from an Indian virus isolate. A/Equine-2/Ludhiana/1/87 (H3N8). Five different preparations of the vaccine were standardized by SRD to prepare 913 doses, each containing 20 micrograms HA/ml-1 dose-1. This test also showed influenza virus subtype specificity as no cross reaction was observed between subtype 1 (H7N7) and subtype 2 (H3N8) viruses.
Subject(s)
Animals , Hemagglutinins, Viral/analysis , Horse Diseases/prevention & control , Horses , Immunodiffusion/methods , Influenza A virus/immunology , Influenza Vaccines/analysis , Orthomyxoviridae Infections/prevention & control , Reference Standards , Vaccines, Inactivated/analysisABSTRACT
Seven hybrid cell lines of mouse myeloma cell line NSO and spleen cells of BALB/c mice producing monoclonal antibodies (MAbs) against equine influenza A/Equi-2/Ludhiana/87 (H3N8) virus were developed. These MAbs were purified, isotyped and characterised by enzyme linked immunosorbent assay (ELISA), fluorescent antibody test (FAT), haemagglutination inhibition (HI) and virus neutralization (VN) tests. The titres of ascitic fluids induced by hybridomas as estimated by ELISA ranged from 1:25,600 to 1:51,200. Monoclonality of these clones was confirmed using a panel of 5 viral antigens, each belonging to a single isotype. MAbs (5) belonged to IgM and one each to IgG1 and IgG2a. Two epitopes appeared to be closely resembling by HI and VN tests but other two epitopes appeared to be different.