摘要
目的:利用HER2/neu胞外配体结合区2(RLD2)从噬菌体抗体库中筛选相应抗体,并进行初步检测。方法:设计合成引物,利用PCR方法克隆出RLD2基因后,将其连接到pET-24a(+)载体中,在大肠杆菌中实现高效表达。对包涵体蛋白经纯化、透析复性后得到目的蛋白。以得到的目的蛋白为靶标,从人源性噬菌体抗体库中进行4轮筛选得到抗体,经ELISA法初步鉴定,并用MTT法检测阳性克隆。结果与结论:初步得到6株亲和力较高的抗HER2/neu抗体,选取其中2株进行了MTT法检测,表明对HER2高表达的乳腺癌细胞有较明显的抑制作用。
Objective： To screen the antibodies of anti-HER2/neu receptor ligand domain 2（RLD2） from the phage library, and identify them by ELISA and MTT. Methods： The gene of RLD2 was amplified by PCR, and cloned into the pET-24a （＋） expression vector and expressed in E.coli. The protein was purified and then renatured by dialysis. Four rounds of selection against RLD2 showed specific enrichment of phage antibody. The antibodies were identified by ELISA and MTT. Results and Conclusion： 6 clones which had higher appetency were obtained. Two clones of them were found that they could inhibit the growth of SKBR-3 breast tumor cells.
出处
《生物技术通讯》
CAS
2006年第4期574-576,共3页
Letters in Biotechnology
