Developments in On-Line Chromatographic Methods Coupled with Mass Spectrometry for the Characterization of Intact Proteins

Library Number:
Part Number:
Paul Rainville, Himanshu Gadgil, Da Ren and Jeff Mazzeo [Waters]
Content Type:
Content Subtype:
Related Products:
LC-MS has become a powerful tool used in the characterization of complex samples. By far the most common mode of chromatography coupled with mass spectrometry has been reversed-phase. The coupling of reversed-phase chromatography with mass spectrometry has been a very effective tool in the characterization of protein pharmaceuticals. The most prominent use of this technique has been peptide mapping, where peptides obtained from proteolytic cleavage of the protein pharmaceutical are separated and identified. The advantage of this technique is the direct identification of peptides as well as modifications that might be present. Modifications such as oxidation, deamidation and glycosylation can all be determined using this technique. Here we show the use of other modes of chromatography coupled with mass spectrometry for the separation, identification, and characterization of intact proteins. Chromatographic separation modes based on size exclusion, ion exchange, reversed-phase, and affinity were evaluated. Each of these chromatographic separation modes coupled with mass spectrometry offers a different dimension in the characterization and separation of intact proteins.

Title Format File Size
wa40509 PDF 454.44kB