Development of anti-pB602L monoclonal antibodies
Six-week-old female mice were immunized three times at a 14 d interval. Three days after the fourth enhanced immunization, the spleen cells of the mice were fused with SP2/0 cells. The cells were cultured in DMEM medium with HTA and 20% FBS. Ten days later, the cell culture supernatant was detected by indirect ELISA (iELISA), and the positive clones were selected and subcloned three times by a limited dilution method. The identified monoclonal cells were cultured in the medium containing 20% FBS and 1% Penicillin Streptomycin. Female mice aged 10 weeks were used to prepare ascites fluid of the monoclonal antibodies. The antibodies titer of ascites was detected by iELISA. SBA Clonotyping System-HRP (Southern Biotech, USA) was used to determine the subclasses of monoclonal antibodies. Western blot and IFA were used to determine the specificity of the monoclonal antibodies against the recombinant protein and virions.