慢性髓细胞白血病BCR-ABL融合基因片段的克隆表达及其单克隆抗体的制备  

Cloning and expression of BCR-ABL fusion gene fragment and preparation of monoclonal antibodies against the recombinant protein

作　　者：张保平[1] 潘兴元[1] 钱莉[1] 晋秋孺[1] 龚卫娟[1] 季明春[1] 

机构地区：[1]扬州大学医学院病原生物学与免疫学教研室,江苏扬州225001

出　　处：《免疫学杂志》2008年第5期549-551,共3页Immunological Journal

基　　金：江苏省自然科学基金重点项目(BK2006706);江苏省自然科学基金项目(BK2004404)

摘　　要：目的制备针对慢性髓细胞白血病(CML)bcr-abl融合蛋白的单克隆抗体(mAb)。方法采用PCR方法扩增包含BCR-ABL(b3a2)融合位点周围约450bp的基因片段,将其定向克隆到pQE-31载体内,转化大肠杆菌M15菌株,诱导表达6×His标签的融合蛋白p18bcr-abl并对其进行纯化,用纯化后的蛋白免疫Balb/c小鼠,采用常规的细胞融合技术制备针对bcr-abl蛋白的mAb并对其特性进行鉴定。结果获得了6株分泌抗bcr-abl融合蛋白mAb的杂交瘤细胞株,所分泌的mAbs均特异性识别bcr-abl蛋白abl端。结论本研究中所表达的融合蛋白p18bcr-abl及其mAb的获得为慢性髓细胞白血病的研究提供了有效的工具。Objective To prepare the monoclonal antibody （mAb） against Chronic myeloid leukemia （CML） bcr-abl fusion protein p18^bcr-abl. Methods The fragment of BCR-ABL fusion gene （b3a2） was amplified by PCR, and then cloned into pQE-31 vector according to the current open reading frame. This recombinant vector was transformed into E. coli M15 and induced by IPTG for expressing 6 × His-tag fusion protein p18^bcr-abl. The protein was purified and used to immunize the Balb/c mice. The specific monoclonal antibodies （mAbs） were prepared by the cell fusion technique. Results Six strains of hybridomas secreting the anti-bcr-abl mAbs were prepared. The mAbs could specifically bind to abl region of the bcr-abl protein. Conclusion The recombinant fusion protein p18^bcr-abl and mAbs will provide effective tools for CML research in our laboratory.

关 键 词：慢性髓细胞白血病 BCR-ABL融合蛋白 克隆表达 单克隆抗体 

分 类 号：R733.72[医药卫生—肿瘤]