抗体文库的噬菌体展示筛选：淘选程序。

Phage Display Selection of Antibody Libraries: Panning Procedures.

作者信息

Peng Haiyong, Rader Christoph

机构信息

Department of Immunology and Microbiology, The Herbert Wertheim UF Scripps Institute for Biomedical Innovation & Technology, University of Florida, Jupiter, Florida 33458, USA.

Department of Immunology and Microbiology, The Herbert Wertheim UF Scripps Institute for Biomedical Innovation & Technology, University of Florida, Jupiter, Florida 33458, USA

出版信息

Cold Spring Harb Protoc. 2024 Oct 15. doi: 10.1101/pdb.prot108602.

DOI:10.1101/pdb.prot108602

PMID:39406533

Abstract

The mining of naive, immune, and synthetic antibody repertoires by phage display has been widely applied to the de novo generation and in vitro evolution of monoclonal antibodies from multiple species. Once built, phage display antibody libraries can be selected by a variety of different strategies tailored toward the desired antigen binding properties. Here, we describe the selection of antibody libraries generated in a phage display vector of the pComb3 phagemid family. The approach includes panning procedures for immobilized antigens, biotinylated antigens in solution, and cell surface antigens. Although the typical format of these antibody libraries is human Fab or chimeric nonhuman/human Fab, the basic selection strategies provided in this protocol are compatible with a variety of formats.

摘要

通过噬菌体展示筛选天然、免疫和合成抗体库已广泛应用于多个物种单克隆抗体的从头生成和体外进化。一旦构建完成，噬菌体展示抗体库可通过针对所需抗原结合特性定制的多种不同策略进行筛选。在此，我们描述了在pComb3噬菌粒家族的噬菌体展示载体中生成的抗体库的筛选方法。该方法包括针对固定化抗原、溶液中的生物素化抗原和细胞表面抗原的淘选程序。尽管这些抗体库的典型形式是人类Fab或嵌合非人类/人类Fab，但本方案中提供的基本筛选策略适用于多种形式。