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目的 构建并筛选小鼠抗人静息 /活化T淋巴细胞表面抗原的噬菌体展示抗体库。方法 以人静息 /活化T淋巴细胞免疫Balb/c小鼠 ,取脾细胞 ,RT -PCR扩增VH 和VK 基因 ,拼接为ScFv ,插入噬菌粒载体转化大肠杆菌 ,构建了噬菌体展示抗体库 ,以人T淋巴细胞为靶抗原 ,对该库进行了 3轮淘洗 ,ELISA方法鉴定噬菌体抗体。结果 成功构建了最大实际库容量为 1 .75× 1 0 6的小鼠抗人静息T淋巴细胞表面抗原的噬菌体展示ScFv抗体库和小鼠抗人活化T淋巴细胞表面抗原的噬菌体展示ScFv抗体库。结论 所构建的抗体库可进一步用于对T淋巴细胞表面蛋白抗原的差异显示分析及构建基因工程抗体。
Objective To construct and screen the phage display antibody library of mouse anti-human resting / activated T lymphocyte surface antigen. Methods Balb / c mice were immunized with human resting / activated T lymphocytes and splenocytes were harvested. VH and VK genes were amplified by RT-PCR and spliced into ScFv. The phagemid vector was inserted into E. coli to construct a phage display antibody library , Human T lymphocytes as target antigen, the library was three rounds of panning, ELISA method to identify phage antibodies. Results The phage display of mouse anti-human resting T lymphocyte surface antigen with the largest actual volume of 1.75 × 106 was successfully constructed. The phage display of ScFv antibody library and mouse anti-human activated T lymphocyte surface antigen showed that ScFv antibody Library. Conclusion The constructed antibody library can be further used to analyze the differential expression of T-lymphocyte surface antigen and to construct engineered antibody.