Selection of human antibody fragments by phage display
Selection of human antibody fragments by phage display
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DOI：
10.1038/nprot.2007.448
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发表时间：
2007-01-01
期刊：
影响因子：
14.8
通讯作者：
Christ, Daniel
中科院分区：
文献类型：
--
作者：
Lee, Carol M. Y.;Iorno, Niccolo;Christ, Daniel
在这里，我们描述了一个协议，用于选择人类抗体片段使用的剧目展示在丝状噬菌体。抗原特异性克隆通过与固定化抗原结合，然后洗脱和噬菌体再增殖来富集。在多轮结合选择后，通过ELISA鉴定特异性克隆。本文综述了噬菌体展示和抗体技术，以及抗原固定化的详细方案，纯化或复合抗原上的库的选择和结合剂的鉴定。
Here, we describe a protocol for the selection of human antibody fragments using repertoires displayed on filamentous bacteriophage. Antigen-specific clones are enriched by binding to immobilized antigen, followed by elution and repropagation of phage. After multiple rounds of binding selection, specific clones are identified by ELISA. This article provides an overview of phage display and antibody technology, as well as detailed protocols for the immobilization of antigen, the selection of repertoires on purified or complex antigens and the identification of binders.