HPLC and MS analysis for the identification and characterisation of peptides presented in the context of the non-classical human leukocytes antigen (HLA) class I molecule HLA-E.
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.
Millo
Enrico
E
Department of Experimental Medicine, Biochemistry section c/o Center of Excellence for Biomedical Research, University of Genoa, Genoa, Italy. enrico.millo@unige.it
Damonte
Gianluca
G
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D000939:Epitopes / Q000276:immunology; D006680:HLA Antigens / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification; D015395:Histocompatibility Antigens Class I / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification; D006801:Humans; D013058:Mass Spectrometry; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry; D022223:Vaccines, Subunit / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification; D000097625:HLA-E Antigens
D000939:Epitopes; D006680:HLA Antigens; D015395:Histocompatibility Antigens Class I; D022223:Vaccines, Subunit; D000097625:HLA-E Antigens