Characterization of major histocompatibility complex-associated peptides from a small volume of whole blood
| Autor: | Joseph F. Leykam, Leann M. Hopkins, Michael Schall, John A. Gerlach |
|---|---|
| Rok vydání: | 2004 |
| Předmět: |
Blood Platelets
chemistry.chemical_classification biology Biophysics Peptide Cell Biology Human leukocyte antigen Mass spectrometry Major histocompatibility complex Biochemistry Mass Spectrometry Amino acid Major Histocompatibility Complex chemistry Liquid chromatography–mass spectrometry MHC class I biology.protein Humans Amino Acids Peptides Molecular Biology Blood Chemical Analysis Chromatography High Pressure Liquid Whole blood |
| Zdroj: | Analytical Biochemistry. 328:155-161 |
| ISSN: | 0003-2697 |
| DOI: | 10.1016/j.ab.2004.02.011 |
| Popis: | Class I major histocompatibility complex (MHC) presents intracellular-derived peptides on the majority of cells within the human body. Intracellular proteins are degraded into peptides of 8–11 amino acids, allowing them to fit into the groove of an empty MHC class I molecule. Detection of MHC-associated peptides can be challenging with the major difficulty being the ability to obtain peptides in adequate concentration. Published protocols require a large sample size that is unrealistic for a clinically available sample. Based on calculations, it should be possible to characterize MHC-associated peptides from cells obtained from 30 ml of whole blood. A citric acid wash of whole platelets was implemented to release the peptides with sample cleanup by reversed-phase high-performance liquid chromotography on a peptide trap. Peptides were analyzed by liquid chromatography tandem mass spectrometry. Four peptides were identified from an individual’s platelets. The binding motifs of the peptides were consistent with the published MHC binding motif of the individual. Since red blood cells do not express MHC, they were used as a negative control. Using citric acid wash of whole cells and a peptide trap, the more abundant MHC-associated peptides can be identified. This report demonstrates the identification of peptides from a sample volume compatible with reasonable clinical availability. |
| Databáze: | OpenAIRE |
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