Study Of Monoclonal Antibodies Against O Type Foot-and-Mouth Disease Virus

The monoclonal antibody (McAb) has strong specific and repeat characters, and it can be make into productions of detection and be used in research fields. The purposes of this paper were for obtain monoclonal antibodies to O type Foot-and-Mouth Disease Virus (FMDV), and for make them into detective productions of FMDV antigen with a rapid, specific, sensitive and convenient mean.By cell culture and virus multiplication methods, the cell culture supernatant which has FMDV was got, and through a series of steps, such as inactivation, concentrate, centrifuge. At last, the purified protein antigen of FMDV was obtained. The BALB/C mice were immunized 4 times with FMDV. The myeloma cell line SP2/0 was fused with the spleen of immunized BALB/C by PEG-1 000, and by several limiting dilution, three specific monoclonal antibodies were screened by indirect ELISA. The three specific monoclonal antibodies cell strains named 2F1, 6H4 and 2G7, respectively. Among the three cell strains, 6H4 and 2G7 belonged to IgG1 subclass of Ig, 2F1 belonged to IgG2b subclass of Ig. The results of specificity cross test indicated that the three cell strains' monoclonal antibodies were not specific reaction with the cell culture supernatant of BHK-21, A, C or Asial type of FMDV. Their titers in cell culture supernatant were 1: 800, 1: 6 400 and 1: 800, respectively. Their titers of abdomen liquor were 1: 1×10~4, 1: 4×10~5 and 1: 1×10~4, respectively.In several passages, the titers of 6H4 abdomen liquor got to 1: 8×10~5, and those of 2F1, 2G7 also have 1: 1×10~4. The purified protein of 6H4 by SDS-PAGE showed the molecular weights of the heavy chain and light chain were about 45.0 kD and 25.0 kD. The number of chromosomal to hybridoma cell strain 6H4 was 100.By mean of ELISA, the antigenic epitopes was detected and recognized by 2F1, 6H4 and 2G7. The results indicated that the epitopes recognized 6H4 overlapped partly that of 2F1, and the epitopes recognized 6H4 is different to that of 2G7.In the appliance research test, lemon acid was made to colloidal gold and marked it with monoclonal antibody 6H4. Monoclonal antibodies of cell strains 6H4 can be make to colloidal gold in pH 8.64 and with 12 mg/L of protein concentration. It also can be make to the colloidal gold marked antibody reaction membrane and hope to take part in to product the detective productions of FMDV antigen.In this whole experiment, we obtained monoclonal antibodies to FMDV, and had a significance trial for make them into detective productions of FMDV antigen in the future.