Trier Nicole Hartwig
Department of Autoimmunology and Biomarkers, Statens Serum Institut, Artillerivej 5, 2300, Copenhagen, Denmark.
Methods Mol Biol. 2015;1348:229-39. doi: 10.1007/978-1-4939-2999-3_20.
Characterization of peptide antibodies through identification of their target epitopes is of utmost importance. Understanding antibody specificity at the amino acid level provides the key to understand the specific interaction between antibodies and their epitopes and their use as research and diagnostic tools as well as therapeutic agents. This chapter describes a straightforward 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: (1) overlapping peptides, used to locate antigenic regions; (2) truncated peptides, used to identify the minimal peptide length required for antibody binding; and (3) 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 fine mapping. The combination of resin-bound peptides and soluble peptides for epitope mapping provides a time-sparing and straightforward approach for characterization of peptide antibodies.
通过鉴定其靶抗原表位来表征肽抗体至关重要。在氨基酸水平上了解抗体特异性是理解抗体与其抗原表位之间特异性相互作用以及将其用作研究和诊断工具以及治疗剂的关键。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抗原表位的直接策略。该方法结合了三种不同类型的肽组以全面表征肽抗体：（1）重叠肽，用于定位抗原区域；（2）截短肽，用于鉴定抗体结合所需的最小肽长度；（3）取代肽，用于鉴定对抗体结合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抗原表位估计，而随后使用可溶性肽进行精细定位。结合树脂结合肽和可溶性肽进行抗原表位定位为肽抗体的表征提供了一种省时且直接的方法。
