关键词: 赤羽病; 赤羽病毒; 单克隆抗体; 制备; 鉴定

Abstract: Balb/c mice were immunized with purified akabane virus, after the fusion of the splenocytes obtained from the immunized mice with SP2/0 cells, we obtained two strains hybridoma cell lines steadily secreting akabane virus specific monoclonal antibodies by indirectELISA screening and three times subcloning, they were named 3A and 2C. ELISA and neutralization test revealed, they were all akabane virus specific monoclonal antibodies, the ELISA titer for akabane virus antibodies in culture supernatant were 1∶640 and 1∶320, the ascites obtained by mice were 1∶256000 and 1∶128000, affinity constant(Ka) were 1.16×10^-9 mol/L and 6.31×10^-8 mol/L, relative affinity of 3A was higher than that of 2C, the titers of the neutralizing antibodies were 1∶64 and 1∶32 respectively. Test for the subgroup of the monoclonal antibodies revealed that 3A and 2C were subgroup IgG1, all their light chains were the kappa type. The two strains hybridoma cell lines can steadily secrete akabane virus specific monoclonal antibodies after three times frozen and resuscitated. It indicates that, we have successfully prepared akabane virus specific McAb, and found an important base for further developing fast diagnostic method to detect akabane virus.

Key words: akabane disease; akabane virus; monoclonal antibody(McAb); preparation; identification