抗人精浆蛋白抗体Fab片段在大肠杆菌中表达及活性鉴定

Prokaryotic expression and bioactivity identification of Fab against human gamma-seminoprotein

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。

AIM: To construct the expression vector containing cDNA encoding Fab against human gamma seminoprotein and express it in E.coli . METHODS: The genes encoding К chain and Fd against gamma seminoprotein were acquired from pUC19 К and pBluescript KS(M13 ) Fd by restrictive enzyme digestion and then cloned into the expression vector pComb3 to construct recombinant expression vector pComb3 Fab. pComb3 Fab was transfected into and expressed in XL1 Blue. RESULTS: Fab against r seminoprotein was expressed in. XL1 Blue. Western blot analysis and immunocytochemical staining demonstrated that expressed Fab could specifically bind to gamma seminoprotein. CONCLUSION: Fab against gamma seminoprotein has been expressed successfully with biological activity, which creat favourable condiction for further study on targeted therapy of prostate cancer.