Department of Neurology, Rigshospitalet Glostrup, Glostrup, Denmark.
Methods Mol Biol. 2024;2821:179-193. doi: 10.1007/978-1-0716-3914-6_14.
Characterization of peptide antibodies through identification of their target epitopes is of utmost importance, as information about epitopes provide important knowledge, among others, for discovery and development of new therapeutics, vaccines, and diagnostics.This chapter describes a strategy for mapping of continuous peptide antibody epitopes using resin-bound and soluble peptides. The approach combines three different types of peptide sets for full characterization of peptide antibodies; (i) overlapping peptides, used to locate antigenic regions; (ii) truncated peptides, used to identify the minimal peptide length required for antibody binding; and (iii) substituted peptides, used to identify the key residues important for antibody binding and to determine the specific contribution of key residues. For initial screening, resin-bound peptides are used for epitope estimation, while soluble peptides subsequently are used for final epitope characterization and identification of critical hot spot residues. The combination of resin-bound peptides and soluble peptides for epitope mapping provides a time-saving and straightforward approach for characterization of antibodies recognizing continuous epitopes, which applies to peptide antibodies and occasionally antibodies directed to larger proteins as well.
通过鉴定肽抗体的靶表位来对其进行特征分析非常重要，因为有关表位的信息为发现和开发新的治疗方法、疫苗和诊断方法提供了重要的知识。本章描述了一种使用树脂结合肽和可溶性肽来绘制连续肽抗体表位的策略。该方法结合了三种不同类型的肽集合，以全面表征肽抗体；（i）重叠肽，用于定位抗原区域；（ii）截断肽，用于确定抗体结合所需的最小肽长度；和（iii）取代肽，用于鉴定对抗体结合很重要的关键残基，并确定关键残基的具体贡献。对于初始筛选，使用树脂结合肽进行表位估计，然后使用可溶性肽进行最终表位特征分析和鉴定关键热点残基。用于表位作图的树脂结合肽和可溶性肽的组合为识别连续表位的抗体的特征分析提供了一种节省时间和直接的方法，适用于肽抗体，偶尔也适用于针对较大蛋白质的抗体。
