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- W2034705509 abstract "The presentation of MHC class I peptides at cell surfaces and the subsequent cytolytic T-lymphocyte response are critical components of the mammalian immune response. However, the identification and sequencing of such peptides present a considerable analytical challenge since > 10 000 peptides at 10−15–10−18 M concentrations are often present in the mixture. We describe a two-dimensional chromatography approach in conjunction with tandem mass spectrometry to sequence and identify such peptides. After immunoaffinity concentration, and subsequent acetic acid release of MHC class I peptides from MHC protein complex, the peptides are subjected to reversed phase HPLC, where they are separated based on their hydrophilic-hydrophobic character. These coarse fractions are then loaded onto a specially designed membrane preconcentration-capillary electrophoresis cartridge (mPC-CE) and subsequently subjected to on-line mPC-CE-MS analysis. The second dimension of chromatography by CE separation affords resolution of peptides based on their charge/mass (to a first approximation) ratio. Ultimately peptides are sequenced using mPC-CE-tandem mass spectrometry (mPC-CE-MS-MS). We describe the strategy for sequencing <60 femtomoles of a peptide obtained from 3·109 Kb-derived EL-4 cells." @default.
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- W2034705509 date "1996-09-01" @default.
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- W2034705509 title "Strategy for isolating and sequencing biologically derived MHC class I peptides" @default.
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- W2034705509 doi "https://doi.org/10.1016/0021-9673(96)00333-0" @default.
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