重复给予恒河猴rhKGF后其免疫原性及抗体中和性质的检测

Immunogenicity and neutralizing activity of the Rhesus monkeys after repeated intravenous injections of rhKGF

目的测定恒河猴经静脉重复给予重组人角质细胞生长因子(rhKGF)后血清中特异性抗体的水平及抗体中和性质。方法采用间接ELISA法检测猴血清中的抗体水平,同时采用32D-KGFR细胞检测抗体的中和活性。结果确定了抗原包被浓度为10μg·mL-1,抗体从1.6×103开始稀释,二抗稀释倍数为1∶5×103的间接ELISA法实验体系;14～56 d均可检测到抗体的存在;生物学活性测定的结果显示其为非中和性抗体。结论重复给予恒河猴rhKGF的毒性试验中,部分猴的血清中检测到抗体,但不是中和抗体。

OBJECTIVE To determine the immunogenicity and neutralizing activity of the Rhesus monkeys after repeated intravenous injections of recombinant human keratinocyte growth factor（rhKGF）.METHODS An indirect ELISA method was established to assay the serum antibody levels in the monkeys,and whether the serum antibodies were neutralizing antibodies was determined by cell-based bioassay.RESULTS The concentration of coated antigen was 10 μg·mL^-1.The antibody dilution started from 1600,1：5×10~3 dilution of indirect ELISA experimental system with secondary antibody was used.The antibody could be detected from 14 d to 56 d.The test result obtained with 32D-KGFR cells showed that the antibody was non-neutralizing antibody.CONCLUSION Antibodies to KGF were detected in the Rhesus monkeys,which had no neutralizing potency.