免疫肽组学分析 HLA Ⅰ类分子展示的磷酸肽组

Immunopeptidomic Analysis of the Phosphopeptidome Displayed by HLA Class I Molecules.

机构信息

Proteomics Unit. Spanish National Biotechnology Centre (CNB-CSIC), Madrid, Spain.

出版信息

Methods Mol Biol. 2022;2420:149-158. doi: 10.1007/978-1-0716-1936-0_12.

DOI:10.1007/978-1-0716-1936-0_12

Abstract

Liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) is a powerful tool that allows the characterization of MHC-associated peptidomes. In addition to conventional nonmodified peptides, these complex mixtures also contain phosphorylated species, which may be of great interest for personalized cancer immunotherapy. Here, we provide a detailed protocol to identify phosphopeptides displayed by human HLA class I molecules consisting of four main steps: (1) immunopurification of MHC-I molecules, (2) phosphopeptide enrichment, (3) LC-MS/MS analysis, and (4) MS/MS ion search.

摘要

液相色谱串联质谱（LC-MS/MS）是一种强大的工具，可用于鉴定 MHC 相关肽组。除了常规的非修饰肽外，这些复杂的混合物还包含磷酸化物质，这对于个性化癌症免疫治疗可能非常重要。在这里，我们提供了一个详细的方案，用于鉴定由人 HLA Ⅰ类分子展示的磷酸肽，该方案包括四个主要步骤：（1）MHC-I 分子的免疫纯化，（2）磷酸肽富集，（3）LC-MS/MS 分析，和（4）MS/MS 离子搜索。