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目的 :建立分泌抗HIV 1p2 4单克隆抗体 (mAb)的杂交瘤细胞株 ,并对其特性进行初步鉴定。方法 :以纯化的基因工程制备的p2 4抗原免疫BALB/c小鼠 ,取免疫小鼠的脾细胞与Sp2 /0骨髓瘤细胞融合 ,经HAT、HT选择培养及有限稀释法进行克隆化后 ,用间接ELISA法及Dotblot对其进行筛选和特性鉴定。结果 :筛选到 2株可分泌抗HIV 1p2 4mAb的杂交瘤细胞 ,其腹水效价为 1×10 -5,亲和力为 1.7× 10 4～ 1.8×10 4mol/L ,mAb的Ig亚类均为IgG1。两株mAb与HBcAg、HCVRNA阳性血清及HIVgp4 1等均无交叉反应 ,只与HIV 1p2 4抗原阳性血清产生特异反应。结论 :成功地建立了 2株可分泌抗HIV 1p2 4mAb的杂交瘤细胞 ,为进一步研制HIV 1p2 4抗原的ELISA检测试剂盒奠定了基础
Objective: To establish a hybridoma cell line secreting anti-HIV 1p24 monoclonal antibody (mAb) and to identify its characteristics. Methods: BALB / c mice were immunized with purified genetically engineered p2 4 antigen. The splenocytes of immunized mice were fused with Sp2 / 0 myeloma cells. After cloning by HAT, HT selective culture and limiting dilution, Indirect ELISA and Dotblot were used to screen and characterize them. Results: Two hybridoma cells secreting anti-HIV 1p2 4 mAb were screened. The ascites titer was 1 × 10 -5 and the affinity was 1.7 × 10 4 ~ 1.8 × 10 4 mol / L. The Ig subclasses of mAb were all IgG1 . The two mAbs did not cross-react with HBcAg, HCVRNA positive serum and HIVgp41, etc., and only reacted specifically with HIV 1p24 antigen positive sera. Conclusion: Two hybridoma cells secreting anti-HIV 1p2 4 mAb were successfully established, which laid the foundation for the further development of an ELISA kit for HIV 1p2 4 antigen