Development And Analysis The Preliminary Application Of Monoclonal Antibody Against Serotype A VP1 Foot-and-mouth Disease Virus

Foot-and-mouth disease (FMD) is a highly contagious communicable disease caused by foot-and-mouth disease virus (FMDV) which always infect cloven-hoofed species and is classified by Office International des Epizooties (OIE) as the first one of OIE List A disease. It has negative influence on politics,economy and society in epidemic countries。The monoclonal antibody (McAb) possess the merit of strong specific,repeat characters and highly stability,so it has been applicated in many fields on FMDV。According to fusing between spleen cells of BALB/c mice aged six weeks immunized with the type A FMDV VP1 and SP2/0 myeloma cells, a series of monoclonal antibodies against serotype A FMDV has been acquired, screened by indirect ELISA and several limiting dilution , 4 strain McAbs have been identified and named 7B11,7H11,8G2,8H4, which have excellent characterization。The McAbs 7B11,7H11,8G2,8H4 titers in cell supernatant and ascites were 1:200,1:200,1:200,1:400 and 1:40960,1:20480,1:20480,1:40960 by the indirect ELISA,respectively。McAb 7B11 and 7H11 belong to IgG1,8H4 and 8G2 IgG2a。In normal situation,myeloma cells posses 62-68 chromosome,spleen cells 40,The result of chromosome analyse was that the average chromosome number of hybridoma cells were between 98 to 108。With the purity of octoic acid and saturated ammonium sulfate,only heavy chain and light chain of antibody,approximately 50KDAAa and 25KDAAa in molecular mass were present in the SDS-PAGE analysis。Indirect immunofluorescence assay (IFA) and enzyme-linked immunsorbent assay (ELISA) indicate that McAbs (7B11,7H11,8G2,8H4) only reacted with serotype A FMDV, but not with O,Asia1 FMDV。Their relative affinity were 8G2>8H4>7H11>7B11 by the indirect ELISA。The additive ELISA showed that the Epitopes of McAbs were same filiform。The latex agglutination text for serotype A FMDV has been established with purified monoclonal antibody,the optimal working dilution of 7B11 and 7H11 were 1:40,and showed highly specificity。We have successfully prepared McAbs against serotype A FMDV, analyzed and identified the biological characterization, based on the foundation of the above,the Latex Agglutination text has been established, it provide valuable experience for the development of diagnostic for the detection of serotype A FMDV。...