Establishment and identification of monoclonal antibody of Golgi protein 73

Objective To establish hybridoma cell line secreting monoclonal antibody (McAb) by Golgi protein 73(GP73) and identify the McAb. Methods Five Balb/c mice were immunized with GP73. The spleen cells of immu-nized mice were fused with sp2/0 cells, screened and cloned. Thus ascites with McAb was prepared. The specificity of GP73 McAb was identified by ELISA and cell immunofluorescence. Results Eleven hybridoma cell lines which could stably secrete specific McAb against GP73 were established with perfect specificity and higher efficiency. After several times of resuscitation and regeneration, the ability of secreting McAb by these eleven cell lines still kept on their specific quality. The specific identifications of GP73 McAb showed positive reaction very well. The concentration of antigen ofGP73 expression of gene engineering was 1 ng/ml by using paired GP73 monoclonal antibody. Conclusion The GP73 McAb secreted by the eleven hybridoma cell lines shows perfect specificity and strong affinity with GP73, which provides the key technical foundation on development of GP73 diagnostic reagent.