抗乙肝病毒表面抗原单链抗体在大肠杆菌中的高效表达

High Level Expression of Anti-HBsAg single-chain Fv in Escherichia coli

在本室构建的抗乙肝病毒表面抗原单链抗体表达载体pHBSCS的基础上,通过PCR将单链抗体基因重组到原核细胞表达载体pT7中,构建了单链抗体高效表达载体pT7SC.将pT7SC质粒转化大肠杆菌BL21(DE3),经IPTG诱导后,获得ScFv的高效表达,经SDS-PAGE检测,表达产物占菌体总蛋白的28%以上.对IPTG诱导后的细菌培养上清进行竞争抑制性ELISA检测,证明所表达的ScFv具有抗体活性.

An anti-HBsAg ScFv expression vector pT7SC was constructed by inserting theScFv gene, which was obtained by PCR amplification from vector pHBSCS construced previouslyin our lab, into a T7 promoter-containing vector pT7. High level expression of ScFv was obtainedby IPTG induction of pT7SC transformed E Coli BL21 (DE3) cells. The ScFv product constituted28N of the total bacterial protein. HBsAg binding activity was also detected in the Supernatant ofthe induced bacteria by competitive ELISA.