抗癌胚抗原单链抗体基因与核心链霉亲和素基因融合在昆虫细胞中的表达

将抗癌胚抗原单链抗体基因与核心链霉亲和素基因融合插入昆虫杆状病毒供体质粒 pFastBacHTa中 ,在粉纹夜蛾Tn 5B1 4细胞中进行表达。SDS PAGE分析结果表明 ,表达产物分子量为 4 1kD左右 ,Western印迹分析结果表明 ,以HRP标记的生物素进行蛋白质印迹在 4 1kD处可见表达条带 ,表明融合蛋白能特异性的与生物素结合 ,放射免疫分析表明重组杆状病毒表达产生的ScFv CS蛋白能特异性结合癌胚抗原

Fused Expression of Anti-CEA Single-chain Antibody Gene with Core-streptavidin Gene in Insect Cells

A fused construct formed by an anti CEA si ng le chain antibody gene with a core streptavidin gene(ScFv CS)was inserted into the donor plasmids pFastBacHTa and expressed in Tn 5B1 4 cells. SDS PAG E analysis revealed that the molecular weight expressed product of ScFv CS were 41 kD. Using horseradish peroxidase(HRP)labeled biotin as antibody product West ern blot, one band of 41 kD only was detected. It was also shown by RIA that the expression product possessed the ability to bind to its specific antigen of CEA . These results showed that the fusion protein not only had the ability to bind CEA, but also could bind biotin specifically.