Biochemical Applications in Proteomics

A major application of HPLC interfaced with MS is in protein or peptide sequencing. This can be accomplished using a wide variety of chromatographic separations followed by different kinds of mass spectrometric characterizations. We will illustrate one example of such techniques to give an impression of the power and complexity of such analyses. A very large protein might consist of 

A method for cutting them specifically and reproducibly into fragments (peptides) that are small enough to he separated easily hy chromatographic techniques (e.g., LC, capillary electrophoresis (CE), 2D-thin-layer chromatography [TLC], electrophoresis) 

A method for interfacing the peptides from the chromatographic separation to some form of spectroscopic detection (primarily MS, due to its superior sensitivity to underivatized peptide ehains or amino acids and the high information content of mass spectra, especially when employed as multistage MS ) 

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