Frenzel André, Kügler Jonas, Wilke Sonja, Schirrmann Thomas, Hust Michael
Abteilung Biotechnologie Technische Universität Braunschweig, Institut für Biochemie, Biotechnologie und Bioinformatik, Braunschweig, Germany.
Methods Mol Biol. 2014;1060:215-43. doi: 10.1007/978-1-62703-586-6_12.
Antibody phage display is the most commonly used in vitro selection technology and has yielded thousands of useful antibodies for research, diagnostics, and therapy.The prerequisite for successful generation and development of human recombinant antibodies using phage display is the construction of a high-quality antibody gene library. Here, we describe the methods for the construction of human immune and naive scFv gene libraries.The success also depends on the panning strategy for the selection of binders from these libraries. In this article, we describe a panning strategy that is high-throughput compatible and allows parallel selection in microtiter plates.
抗体噬菌体展示是最常用的体外筛选技术,已产生了数千种用于研究、诊断和治疗的有用抗体。利用噬菌体展示成功生成和开发人重组抗体的前提是构建高质量的抗体基因文库。在此,我们描述构建人免疫和天然单链抗体基因文库的方法。成功还取决于从这些文库中筛选结合物的淘选策略。在本文中,我们描述了一种与高通量兼容且允许在微孔板中进行平行筛选的淘选策略。