利用大型天然噬菌体抗体库筛选抗死亡受体5人源抗体

发布时间：2018-01-19 23:08

本文关键词： 抗死亡受体抗体人源单链抗体大容量噬菌体抗体库　出处：《细胞与分子免疫学杂志》2015年06期 　论文类型：期刊论文

【摘要】：目的从大容量天然噬菌体抗体库中筛选抗人死亡受体5(DR5)单链抗体(sc Fv)并对其抗原结合活性进行鉴定。方法以前期真核表达纯化的DR5-Fc蛋白包被筛选管,从自主构建的大容量天然噬菌体抗体库中,通过4轮筛选过程获得噬菌体抗体,采用免疫细胞化学染色、ELISA鉴定其抗原结合活性;DNA指纹图谱分析和序列测定确定其抗体类型。结果经过4轮筛选,获得26株与DR5特异性结合的阳性克隆,DNA指纹分析有4种不同的可变区片段;经过基因测序,分析可变区基因的亚群,并将其制备成可溶性的抗体,进一步用免疫细胞化学技术证实所获得的抗体可特异性结合SW480细胞的DR5蛋白。结论利用噬菌体抗体库技术成功获得了特异性抗DR5的人源性sc Fv。
[Abstract]:Objective to screen the anti-human death receptor 5 DR5 single chain antibody (sc FV) from a large volume natural phage antibody library. The antigen-binding activity was identified. Methods the purified DR5-Fc protein was coated with a screening tube. Phage antibodies were obtained from a large volume natural phage antibody library constructed by ourselves and their antigen-binding activities were determined by immunocytochemical staining Elisa. Results after four rounds of screening, 26 positive clones with specific binding to DR5 had four different variable regions. After gene sequencing, the subpopulations of variable region genes were analyzed, and the soluble antibodies were prepared. It was further confirmed by immunocytochemistry that the obtained antibody could specifically bind to the DR5 protein of SW480 cells. Conclusion the phage antibody library technique has been successfully used to obtain the specific anti-#en2# human sc. Fv.
【作者单位】：第二军医大学海军临床医学院普通外科;海军总医院中心实验科;
【基金】：国家科学自然基金(31470897,81172119)
【分类号】：R392
【正文快照】： 死亡受体5(death receptor 5,DR5)[1]属于肿瘤坏死因子受体超家族成员,主要分布于肺癌、卵巢癌、胰腺癌、肝癌、结直肠癌、前列腺癌、皮肤癌等多种肿瘤细胞的表面,在正常细胞很少或几乎不表达,当DR5与其配体肿瘤坏死因子相关凋亡诱导配体(tumor necrosis factor-related apopt

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