Study On Preparation And Application Of Monoclonal Antibodies Against Type Asia1 FMDV

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 Asial 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.The BALB/C mice were immunized 4 times with Asial type FMDV. VP1 protein。The myeloma cell line SP2/0 was fused with the spleen of immunized BALB/C by PEG-1000, and by several limiting dilution, three specific monoclonal antibodies were screened by indirect ELISA. The three specific monoclonal antibodies cell strains named 1B8, 5E1 and 5E2, respectively.Among the three cell strains, 1B8 belonged to IgG₁ subclass of Ig, 5E1 and 5E2 belonged to IgG2b and IgG2a subclass of Ig,. The results of specificity cross test indicated that the three cell strains' monoclonal antibodies were not specific reaction with the A, C or O type of FMDV. Their titers in cell culture supematant were 1∶10⁴, Their titers of abdomen liquor were 1∶4×10⁶, respectively.The purified protein of monoclon 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 SP2/0 cell strains were 50, to hybridoma cell strains were 104。By mean of indirect ELISA, the antigenic epitopes was detected and recognized by1B8, 5E1 and 5E2. The results indicated that the epitopes recognized 1B8 overlapped partly that of 5E2, and the epitopes recognized 1B8 is different to that of 5E1.In the appliance research test, lemon acid was made to colloidal gold and marked it with monoclonal antibody. The strips can recognize Asial type FMDV and VP1 protein, can detective 10^-4 TCID₅₀ virus particle, can store three months in different temperature condition。through experiment we find these strips have highly specificity,sensitivity and stability。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.