利用噬菌体抗体库技术制备抗甲状旁腺激素相关蛋白Fab小分子抗体

Generation of monoclonal antibody Fab fragments to parathyroid hormone related protein by phage display technology

目的 利用噬菌体抗体库技术制备抗甲状旁腺激素相关蛋白 (PTHrP)Fab抗体。方法从经PTHrP免疫的Balb/C小鼠脾细胞提取总RNA ,用逆转录 聚合酶链反应 (RT -PCR)扩增一组Ⅰg轻链和重链Fab段基因 ,插入噬菌体载体pComb3,构建了抗PTHrPFab抗体基因库。用合成短肽和重组PTHrP对抗体库进行了富集筛选 ,获得鼠源抗PTHrPFab段基因后 ,进一步在大肠埃希菌表达 ,并对表达产物的特异性作了鉴定。结果 成功构建了抗PTHrP抗体基因库 ,且筛选到PTHrP特异的鼠源单抗Fab段基因。结论 所获得的Fab抗体能特异性地识别PTHrP抗原。

Objective To develop monoclonal antibody Fab fragments using display technique.Methods The mouse IgG Fab genes of heavy and light chains were amplified from spleen cells of a parathyroid hormone relaed protein(PTHrP) immunized mouse.The combinatorial phage antibody library was prepared by inserting both heavy and light chain Fab genes into phagemid vector pComb3 and followed by infection of helper phage. The library was selected by purified recombinant PTHrP.Results The combinatorial phage antibody library was constructed successfully and the specific mouse Fabs to PTHrP were delected and expressed in E.coli.Conclusion The selected specific mouse Fabs can recognize PTHrP with high specificity.