Department of Experimental Medicine, Biochemistry section c/o Center of Excellence for Biomedical Research, University of Genoa, Genoa, Italy.
Curr Pharm Des. 2009;15(28):3325-35. doi: 10.2174/138161209789105126.
In addition to a variety of other techniques used in T-cell epitope identification, mass spectrometery coupled to liquid chromatography have now become an important and sensitive tool in separation, detection, and sequence analysis of highly complex natural major histocompatibility complex (MHC) ligand mixtures. In this article, we present current strategies for the identification of MHC eluted peptides using high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS) with a particular recall to those presented in the context of the non classical human leukocytes antigen (HLA) class I molecule HLA-E. In addition we also discuss the advantages and disadvantages of the methods available in the literature to concentrate and fractionate the peptides prior to analysis by mass spectrometry. An application of our method for isolation and characterization of peptides presented in the context of HLA-E is finally reported.
除了用于 T 细胞表位鉴定的各种其他技术外,质谱法与液相色谱法相结合现在已成为分离、检测和高度复杂的天然主要组织相容性复合物 (MHC) 配体混合物的序列分析的重要和敏感工具。在本文中,我们介绍了使用高效液相色谱法与串联质谱法 (HPLC-MS/MS) 鉴定 MHC 洗脱肽的当前策略,特别提到了在非经典人类白细胞抗原 (HLA) 类 I 分子 HLA-E 背景下提出的那些策略。此外,我们还讨论了文献中可用于在进行质谱分析之前浓缩和分级肽的方法的优缺点。最后报告了我们的方法在 HLA-E 背景下分离和鉴定肽的应用。