特异性抗IBDV单链抗体基因的构建及序列分析

从抗 IBDV杂交瘤细胞系 SJS中分离总 RNA,经 RT-PCR体外扩增重、轻链可变区基因 ,通过一连接肽 (Giy4 Ser) 3基因拚接形成 Sc Fv基因 ,经 IBDV抗原亲和筛选出特异性阳性克隆进行核苷酸序列测定及氨基酸序列推导。所获得的特异性抗 IBDV单链抗体基因为 VH-Linker-VL 结构 ,其含有编码 (Giy4 Ser) 3的连接肽基因及维持抗体结构所必须的半胱氨酸残基 ,编码产物具有与亲本抗体相同的抗原结合活性和特异性 ,从而为研制具有应用潜力的高表达工程化抗体奠定了基础。

Construction and sequencing of variable region genes of the single strand antibody recognizing IBDV

To obtain the genes of a single chain antibody from mAb SJS against IBDV,The gene of variable region of heavy and light chain were amplified separately by RT PCR from the hybridoma cell line secreting mAb SJS and assembledinto ScFv gene with a (Gly 4Ser) 3 linker by the splicing overlap extension(SOE).The IBDV specific ScFv gene was sequenced after affinitive selection.The results showed that the fusion gene which was capable of encoding affinitive and specific product against IBDV consist of the variable regions of light and heavy chain in mouse anti IBDV antibody and a 45bp Linker .The constructed ScFv gene is a functional ScFv of the specific mAb SJS against IBDV.