目的 :表达具有中和活性的抗基孔肯亚病毒人 鼠嵌合抗体。方法 :用RT PCR法克隆具有中和活性的抗基孔肯亚病毒鼠源单克隆抗体CHIK IF1的轻重链可变区基因 ,克隆人IgG1恒定区基因组基因 ,构建了轻重链嵌合基因和表达质粒pCdhfr1L ,pcDNA3.1H ,共转染二氢叶酸还原酶缺陷型CHO细胞 ,用ELISA检测培养上清中嵌合抗体的表达 ,MTX加压筛选表达量高的克隆 ,扩大培养收集上清并纯化 ,纯化的嵌合抗体进行SDS PAGE、Western印迹检测和抗原结合活性的检测。结果与结论 :在CHO细胞中成功表达了抗基孔肯亚病毒的嵌合抗体 ,为制备抗基孔肯亚病的被动免疫制剂奠定了良好的基础。 OBJECTIVE: To express chimeric antibodies against Chikungunya virus with neutralizing activity. Methods: The light chain variable region (CHV) variable region of murine monoclonal antibody against chikungunya virus CHIK IF1 with neutralizing activity was cloned by RT-PCR. The gene of human IgG1 constant region was cloned and the light and heavy chain chimeric genes were expressed and expressed Plasmids pCdhfr1L and pcDNA3.1H were co-transfected with CHO cells deficient in dihydrofolate reductase. The expression of chimeric antibody in the culture supernatant was detected by ELISA. The clones with high expression level were screened by MTX and the supernatant was collected and purified The purified chimeric antibody was tested for SDS PAGE, Western blot and antigen binding activity. RESULTS AND CONCLUSION: The chimeric antibody against Chikungunya virus was successfully expressed in CHO cells, which laid a good foundation for the preparation of passive immunization against Chikungunya.