基于抗原表位制备抗人GP73单克隆抗体

Generation of monoclonal antibody to GP73 based on antigen epitope

目的：制备抗GP73蛋白的单克隆抗体（mAb）。方法：将GW／3蛋白N末端的肽段（AAAERGAVELK）展示于T7噬菌体表面，扩增重组噬菌体作为抗原免疫小鼠，制备抗体。通过ELISA法检测小鼠血清抗体的效价，筛选分泌抗GP73蛋白抗体的杂交瘤细胞株；通过ELISA、Western blot等方法检测mAb的特异性。结果：利用表面展示GP73抗原表位的重组噬菌体为抗原免疫小鼠，通过细胞融合和筛选得到了能识别GP73蛋白的mAb，且特异性较好。结论：将蛋白质抗原的合适抗原表位展示在T7噬菌体表面，用这种重组噬菌体作为替代抗原可以制备识别全蛋白的mAb。

AIM： Preparation of monoclonal antibody （mAb） against GP73 protein. METHODS： The N-terminal peptide （AAAERGAVELK） of GP73 protein was displayed on T7 phage, the recombinant phage was amplified and used as the immunogen to immunize mouse to produce anti- body. The titer of the antiserum and the positive hybridoma clones which secreted the mAb against GP73 protein were detected by ELISA. The mAb specificity was assayed by ELISA and Western blot. RESULTS： The high specificity mAb against GP73 protein was selected from the mouse im- munized with the recombinant T7 phages displaying the epitope of GP73 by cell fusion and screening. CONCLU- SION： The appropriate protein epitope displayed on T7 phage could be used as alternative antigen to immunize animals to make specific antibody against the corresponding native protein.