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目的 :构建编码抗人精浆蛋白抗体Fab基因的表达载体 ,并在大肠杆菌中进行表达。方法 :从克隆载体pUC19 К和pBluescriptKS(M13 ) Fd中 ,酶切获得抗人精浆蛋白单克隆抗体 (mAb)Fd基因和К链基因。然后将Fd和К链基因重组到Fab表达载体pComb3中 ,构建抗人精浆蛋白Fab基因的重组表达载体pComb3 Fab ,并在XL1 Blue菌中表达。结果 :经重组表达载体转化的XL1 Blue菌株可表达Fab基因。Westernblot和免疫细胞化学分析表明 ,表达产物Fab具有特异性结合精浆蛋白的活性。结论 :抗人精浆蛋白Fab基因成功地获得 ,为进一步将其与抗肿瘤药物偶联用于前列腺癌的导向治疗创造了条件。表达为构建其它基因工程抗体提供了基础。
OBJECTIVE: To construct an expression vector encoding the Fab gene of anti-human seminoprotein antibody and express it in E. coli. Methods: Fd gene and К chain gene of monoclonal antibody (mAb) against human seminal plasma protein were obtained by digestion with cloning vector pUC19 К and pBluescript KS (M13) Fd. The Fd and К chain genes were then recombined into the Fab expression vector pComb3 to construct a recombinant expression vector pComb3 Fab for the anti-human seminoprotein Fab gene and expressed in XL1 Blue bacteria. Results: The XL1 Blue strain transformed with recombinant expression vector can express Fab gene. Western blot and immunocytochemical analysis showed that the expressed product Fab had the activity of binding to the seminoprotein specifically. CONCLUSION: The anti-human seminoprotein Fab gene was successfully obtained, which provided the conditions for its further antitumor drug conjugation for the treatment of prostate cancer. Expression provides the basis for the construction of other genetically engineered antibodies.