抗-HEV单克隆抗体B4C重链可变区基因的克隆及初步表达

Cloning and expression of a heavy chain variable region of mAb B4C against HEV

抗-HEV87A株单克隆抗体（mAb）B4C经体外研究鉴定，证实对我国暴发和散发性HEV毒株有很高的特异性。为从分子水平上了解mAbB4C，我们利用基因工程技术，从杂交瘤细胞系中克隆出了重链可变区基因，并对其全部核苷酸序列进行了分析，可能属于小鼠免疫球蛋白重链IA亚组。将VHcDNA克隆到高效表达载体pQE－31中，经IPTG诱导，SDS－PAGE分析，可见1条Mr为16000左右的表达带，表达产物占菌体总蛋白的29．3％，主要以包涵体形式存在。

The monocloed antibody B4C aghnst HEV strain 87A was Puned to be SPecific to chinese styains Of the epidemic and sporadic HEV. The gene of heaVy chain vallable 'don was amp~ fad the hybridoma Cells and cloned for mpuence analysis. It appeared tO be in the subgroup lA Of mouse edunoglobulin VH. Wad the VH gene was cloned intO a highly expressive vector poE-31 and induced by IPTG, a recolnbinant protein about 16 000(Mr) was detected by SDS-PACE. The yield was up to 29. 3% Of the total amount Of bacteria protein and the pnduct was mostly in the form Of incluaion boas.