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.
出处
《免疫学杂志》
CAS
CSCD
北大核心
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)
