周质间隙
大肠杆菌
抗体
抗原
操纵子
紫胶操纵子
流式细胞术
分子生物学
炭疽杆菌
生物
细菌外膜
化学
免疫球蛋白轻链
基因
细菌
生物化学
遗传学
作者
Yariv Mazor,Thomas Van Blarcom,Brent L. Iverson,George Georgiou
出处
期刊:Methods in molecular biology
日期:2008-12-10
卷期号:: 217-239
被引量:12
标识
DOI:10.1007/978-1-59745-554-1_11
摘要
We have developed a technology for the facile isolation of full-length IgG antibodies with desired specificity from combinatorial libraries expressed in Escherichia coli. Full-length heavy and light chains are expressed from a bicistronic operon and are secreted into the periplasm where they assemble into aglycosylated IgGs that are fully functional for antigen binding. Expression of an inner membrane-tethered Fc-binding protein is used to capture the IgG molecules and anchor them to the cell. Following outer membrane disruption, clones expressing IgGs that specifically recognize fluorescently labeled antigen are selected by flow cytometry. This technique was used for the isolation of several IgGs with nanomolar affinities toward the protective antigen of Bacillus anthracis from immune libraries. High-throughput isolation of E. coli-derived full-length IgG can greatly expedite the discovery and production of antibodies for therapeutic and diagnostic applications.
科研通智能强力驱动
Strongly Powered by AbleSci AI