Garvan Institute of Medical Research, Darlinghurst, Sydney, New South Wales, Australia.
Nat Protoc. 2012 Feb 2;7(2):364-73. doi: 10.1038/nprot.2011.448.
Here we describe protocols for the expression of human antibody fragments in Escherichia coli. Antigen-specific clones are identified by soluble fragment ELISA and concentrated by periplasmic preparation. They are then further purified by affinity chromatography. This article provides an overview of expression and purification strategies for human antibody fragments, as well as detailed protocols for the identification of high-affinity binders and for affinity maturation.
在这里，我们描述了在大肠杆菌中表达人抗体片段的方案。通过可溶性片段 ELISA 鉴定抗原特异性克隆，并通过周质制备进行浓缩。然后，它们通过亲和层析进一步纯化。本文提供了人抗体片段表达和纯化策略的概述，以及鉴定高亲和力结合物和亲和力成熟的详细方案。
