Preparation And Application Of Monoclonal Antibody(MAb) Against Non-structural Protein 3B Of Foot-and-Mouth Disease Virus

The monoclonal antibodies against protein 3B were generated by fusion spleen cells from immunized BALB/c mouse with SP2/0 mouse myeloma cells. The culture of hybridoma lines of first screened by I-ELISA for detection of specific mouse IgG against FMDV 3B protein. Six strain of hybridoma which secreted monoclonal antibodies against protein 3B were determined and given the name 3E5, 4B1, 4D7, 4E11, 7B2 and 8B11. The isotypes were identified to be IgG1 and IgG2b. The titer of these MAbs were over 105 by indirect ELISA. Enzyme-linked immunsorbent assay (ELISA) showed a high specificity to protein 3B and had a low reactivity to FMDV structure proteins and other non-structure protein, revealing that they are specific to FMDV NSP 3B. The result of additive ELISA showed that they could recognize the same antigen epitopes. The MAbs showed strong reactivity in IFA for detection of the FMDV infected BHK-21cell. These hybridoma cells could be cultured for continuous three months and kept a stable condition.A liquid-phase blocking ELISA for the fixed quantity detection of FMDV 3B was developed by using expressed protein 3B as capture antigen and monoclonal antibody 4B1 as detector antibody. With this assay the FMDV NSP 3B was detected at a concentration of 5ng/mL.Using the method of detection of FMDV of different batches of virus antigen in cell culture samples.The results showed that different batches of samples 3B protein content are different, but also before and after inactivation are varies. The establishment of this method for quality control of FMD vaccine antigen has provided quality control for reference.