Chan Yee-Peng, Yan Lianying, Feng Yan-Ru, Broder Christopher C
Uniformed Services, University of the Health Sciences, Bethesda, MD, USA.
Methods Mol Biol. 2009;525:31-58, xiii. doi: 10.1007/978-1-59745-554-1_2.
Neutralizing antibodies are a critical component in the protection or recovery from viral infections. In the absence of available vaccines or antiviral drugs for many important human viral pathogens, the identification and characterization of new human monoclonal antibodies (hmAbs) that are able to neutralize viruses offers the possibility for effective pre- and/or post-exposure therapeutic modalities. Such hmAbs may also help in our understanding of the virus entry process, the mechanisms of virus neutralization, and in the eventual development of specific entry inhibitors, vaccines, and research tools. The majority of the more recently developed antiviral hmAbs have come from the use of antibody phage-display technologies using both naïve and immune libraries. Many of these agents are also enveloped viruses possessing important neutralizing determinants within their membrane-anchored envelope glycoproteins, and the use of recombinant, soluble versions of these viral glycoproteins is often critical in the isolation and development of antiviral hmAbs. This chapter will detail several methods that have been successfully employed to produce, purify, and characterize soluble and secreted versions of several viral envelope glycoproteins which have been successfully used as antigens to capture and isolate human phage-displayed monoclonal antibodies.
中和抗体是病毒感染防护或康复过程中的关键组成部分。对于许多重要的人类病毒病原体，在缺乏可用疫苗或抗病毒药物的情况下，鉴定和表征能够中和病毒的新型人单克隆抗体（hmAb）为有效的暴露前和/或暴露后治疗方式提供了可能。此类hmAb也有助于我们理解病毒进入过程、病毒中和机制，以及最终开发特异性进入抑制剂、疫苗和研究工具。最近开发的大多数抗病毒hmAb来自使用天然和免疫文库的抗体噬菌体展示技术。其中许多病原体也是包膜病毒，在其膜锚定包膜糖蛋白中具有重要的中和决定簇，使用这些病毒糖蛋白的重组可溶性形式通常对于抗病毒hmAb的分离和开发至关重要。本章将详细介绍几种已成功用于生产、纯化和表征几种病毒包膜糖蛋白的可溶性和分泌形式的方法，这些糖蛋白已成功用作抗原以捕获和分离人噬菌体展示单克隆抗体。
