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.