Development and Characterization of Mouse Anti-canine PD-1 Monoclonal Antibodies
In order to screen the monoclonal antibody against canine programmed death receptor 1(cPD-1),BALB/c mice were immunized with prokaryotically expressed and purified recombinant cPD-1 extracellular region protein as the target antigen,and a hybridoma cell line 1F9,which could stably secrete anti-cPD-1 monoclonal antibody,was obtained after 3 rounds of screening and clonation by applying monoclonal antibody hybridoma cell technology and indirect ELISA method.Indirect ELISA showed that the 1F9 hybridoma cell line was able to stably secrete monoclonal antibodies with a potency of 1∶2048 in continuous passaging culture,and the heavy chain was identified as IgG1 and the light chain as κ by subtype identification;the cPD-1 extracellular region protein expressed by HEK293 cells was used as the detection antigen,and Western-blot and indirect immunofluorescence assays were used for identification;the results showed that 1F9 monoclonal antibody could specifically bind to cPD-1 extracellular region protein.In this study,a mouse-derived cPD-1 monoclonal antibody was successfully screened by mouse hybridoma technique.
NETL
NSTL
万方数据
