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将抗癌胚抗原嵌合重链抗体与核心链霉亲和素融合基因插入昆虫杆状病毒供体质粒pFastBacHTa中 ,经大肠杆菌DH10Bac体内转座 ,产生重组杆状病毒pBacHTa VH Cr3 CS。将其转染粉纹夜蛾Tn 5B1 4细胞 ,经扩增后在细胞内进行表达。SDS PAGE分析结果表明 ,在粉纹夜蛾Tn 5B1 4细胞中都表达产生一条特异性蛋白质 ,其相对分子质量约为 75 0 0 0左右 ,以Westernblot分析表明 ,该特异条带即为VH Cr3 CS蛋白。SDS PAGE和Westernblot分析结果表明 ,以HRP标记的生物素作为抗体进行蛋白质印迹在相对分子质量 75 0 0 0处可见表达条带 ,表明融合蛋白能特异性的与生物素结合 ,RIA表明重组杆状病毒表达产生的VH Cr3 CS蛋白能与CEA有较高的结合力。
The anti-carcinoembryonic antigen chimeric heavy chain antibody and the core streptavidin fusion gene were inserted into the insect baculovirus donor plasmid pFastBacHTa and transposed in vivo by E. coli DH10Bac to generate the recombinant baculovirus pBacHTaVHCr3CS. It was transfected with Tn 5B1 4 cells of C. zea. After amplification, it was expressed in cells. The results of SDS PAGE analysis showed that a specific protein was expressed in Tn 5B14 cells. The relative molecular weight was approximately 7500. Western blot analysis showed that the specific band was VH Cr3 CS. protein. The results of SDS PAGE and Western blot analysis showed that HRP-labeled biotin was used as an antibody for Western blotting. The expression band was observed at a molecular weight of 750 000, indicating that the fusion protein could specifically bind to biotin, and RIA showed that the recombinant rod was VH Cr3 CS protein produced by virus expression can have high binding affinity with CEA.