噬菌体展示技术在微孔板中进行抗体选择。

Antibody Selection via Phage Display in Microtiter Plates.

机构信息

Institut für Biochemie, Biotechnologie und Bioinformatik, Departments Biotechnology and Medical Biotechnology, Technische Universität Braunschweig, Braunschweig, Germany.

Abcalis GmbH, Braunschweig, Germany.

出版信息

Methods Mol Biol. 2023;2702:247-260. doi: 10.1007/978-1-0716-3381-6_12.

DOI:10.1007/978-1-0716-3381-6_12

PMID:37679623

Abstract

The most common and robust in vitro technology to generate monoclonal human antibodies is phage display. This technology is a widely used and powerful key technology for recombinant antibody selection. Phage display-derived antibodies are used as research tools, in diagnostic assays, and by 2022, 14 phage display-derived therapeutic antibodies were approved. In this review, we describe a fast high-throughput antibody (scFv) selection procedure in 96-well microtiter plates. The given detailed protocol allows the antibody selection ("panning"), screening, and identification of monoclonal antibodies in less than 2 weeks. Furthermore, we describe an on-rate panning approach for the selection of monoclonal antibodies with fast on-rates.

摘要

最常见和最强大的体外技术来产生单克隆人抗体是噬菌体展示。这项技术是一种广泛应用的、强大的重组抗体选择的关键技术。噬菌体展示衍生的抗体被用作研究工具，在诊断测定中，并在 2022 年，有 14 种噬菌体展示衍生的治疗性抗体获得批准。在这篇综述中，我们描述了一种快速的高通量抗体（scFv）在 96 孔微量滴定板中的选择程序。给定的详细方案允许在不到 2 周的时间内进行抗体选择（“淘选”）、筛选和单克隆抗体的鉴定。此外，我们还描述了一种用于选择具有快速结合速率的单克隆抗体的结合速率淘选方法。