通过免疫亲和纯化分离主要组织相容性复合体（MHC）相关肽段

Isolation of Major Histocompatibility Complex (MHC)-Associated Peptides by Immunoaffinity Purification.

作者信息

Penny Sarah A, Malaker Stacy A

机构信息

Institute of Immunology and Immunotherapy, University of Birmingham, Birmingham, UK.

Department of Chemistry, Stanford University, Stanford, CA, USA.

出版信息

Methods Mol Biol. 2019;2024:235-243. doi: 10.1007/978-1-4939-9597-4_14.

DOI:10.1007/978-1-4939-9597-4_14

PMID:31364053

Abstract

MHC molecules on the surface of cells are responsible for the presentation of antigenic peptides to CD8+ and CD4+ T cells. Downstream analysis of these peptides can offer insight into various disease processes with an immune component, as in autoimmune diseases and cancer. A critical step in studying MHC-associated peptides is their isolation from MHC molecules on the surface of cells. In this chapter, we detail anti-MHC antibody conjugation to beads, immunoaffinity purification of the MHC molecules, and peptide elution. This method can be used for analysis of unmodified and/or posttranslationally modified peptides by mass spectrometry.

摘要

细胞表面的主要组织相容性复合体（MHC）分子负责将抗原肽呈递给CD8 +和CD4 + T细胞。对这些肽进行下游分析可以深入了解各种具有免疫成分的疾病过程，如自身免疫性疾病和癌症。研究与MHC相关肽的关键步骤是将它们从细胞表面的MHC分子中分离出来。在本章中，我们详细介绍了抗MHC抗体与磁珠的偶联、MHC分子的免疫亲和纯化以及肽的洗脱。该方法可用于通过质谱分析未修饰和/或翻译后修饰的肽。