Reineke Ulrich
Lead Discovery Biology Department, Jerini AG, Invalidenstr. 130, D-10115 Berlin, Germany.
Methods Mol Biol. 2009;524:203-11. doi: 10.1007/978-1-59745-450-6_14.
Identification of antibody binding peptides may be based on the primary structure of the protein antigens used to raise the antibodies (knowledge- or sequence-based approach). This involves scanning the entire sequence of the antigen with overlapping peptides (peptide scan), which are then probed for binding to the respective antibody. If a natural protein binding partner is not known, one has to use combinatorial synthetic libraries with peptide mixtures, randomly generated chemically synthesized libraries of single individual sequences, or biologically produced libraries (e.g., phage display libraries, see Chapter "Epitope Mapping Using Phage Display Peptide Libraries"). This chapter describes chemically synthesized combinatorial, as well as randomly generated peptide libraries, collectively called de novo approaches, and their application for antibody epitope mapping.
抗体结合肽的鉴定可基于用于产生抗体的蛋白质抗原的一级结构(基于知识或序列的方法)。这涉及用重叠肽扫描抗原的整个序列(肽扫描),然后检测这些肽与相应抗体的结合情况。如果天然的蛋白质结合伴侣未知,则必须使用含有肽混合物的组合合成文库、随机生成的单个序列化学合成文库或生物产生的文库(例如,噬菌体展示文库,见“使用噬菌体展示肽文库进行表位作图”一章)。本章描述化学合成的组合文库以及随机生成的肽文库,统称为从头合成方法,及其在抗体表位作图中的应用。