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目的获得生物活性更高的癌胚抗原 (CEA)微型抗体用于放免显像。 方法将抗癌胚抗原微型抗体基因插入昆虫杆状病毒供体质粒pFastBacHTb中 ,经大肠杆菌DH10Bac体内转座 ,产生重组杆状病毒BacHT -VH-L ,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。 结果SDS -PAGE分析结果表明 ,在Tn- 5B1- 4细胞中表达产生一条特异性蛋白质 ,其分子量为 2 6kd左右 ;以Westernblot分析表明 ,该特异条带即为VH-L蛋白。RIA表明重组杆状病毒表达产生的VH -L蛋白能特异性的结合CEA ,较大肠杆菌表达物活性更高。结论昆虫细胞表达可制备生物活性更高的癌胚抗原微型抗体。
Objective To obtain a more biologically active carcinoembryonic antigen (CEA) minibody for radioimmunoassay. Methods The anti-carcinoembryonic antigen minibody gene was inserted into the insect baculovirus donor plasmid pFastBacHTb and transfected with E. coli DH10Bac in vivo to generate the recombinant baculovirus BacHT-VH-L, which was transfected with the wormworm (Tn - 5B1-4 cells, which are expressed in cells after amplification. Results SDS-PAGE analysis showed that a specific protein was expressed in Tn-5B1-4 cells with a molecular mass of approximately 26kd. Western blot analysis showed that the specific band was VH-L protein. RIA showed that VH-L protein produced by recombinant baculovirus expression specifically binds to CEA and is more active than E. coli expression. Conclusion The expression of insect cells can be used to prepare carcinoembryonic antigen minibody with higher biological activity.