Phage Display - Methods and Protocols
Phage Display - Methods and Protocols
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噬菌体展示 - 方法和方案
DOI：
10.1007/978-1-0716-3381-6_25
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Tsoumpeli M
中科院分区：
文献类型：
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作者：
Tsoumpeli M
肽噬菌体展示在历史上已用于表位作图单克隆抗体。最近，通过将该方法与下一代测序（所谓的下一代噬菌体展示，NGPD）结合以大量筛选肽结合事件，该方法已成功地应用于绘制对感染的多克隆抗体应答。这导致代表病原体表位的模拟表位组的鉴定。使用这种方法的一个潜在优点是，模拟表位不仅可以代表线性表位，还可以代表构象表位或由蛋白质的翻译后修饰或由其他非蛋白质大分子产生的那些表位。病原体的这种复杂的免疫识别的映射可以为新的血清学测定开发和疫苗设计提供信息。在这里，我们提供了详细的方法应用NGPD来确定面板的mimotopes，特异性识别的抗体从个人与特定的感染。
Peptide phage display has historically been used to epitope map monoclonal antibodies. More recently, by coupling this method with next-generation sequencing (so-called next-generation phage display, NGPD) to mass screen peptide binding events, the methodology has been successfully applied to map polyclonal antibody responses to infection. This leads to the identification of panels of mimotopes that represent the pathogen’s epitopes. One potential advantage of using such an approach is that the mimotopes can represent not just linear epitopes but also conformational epitopes or those produced from post-translational modifications of proteins or from other non-protein macromolecules. The mapping of such complex immunological recognition of a pathogen can inform novel serological assay development and vaccine design. Here, we provide detailed methods for the application of NGPD to identify panels of mimotopes that are recognized specifically by antibodies from individuals with a particular infection.