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Reverse-phase capillary high performance liquid chromatography/high performance electrospray ionization mass spectrometry: an essential tool for the characterization of complex glycoprotein digests

โœ Scribed by Katalin F. Medzihradszky; Marc J. Besman; A. L. Burlingame


Publisher
John Wiley and Sons
Year
1998
Tongue
English
Weight
226 KB
Volume
12
Category
Article
ISSN
0951-4198

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โœฆ Synopsis


The B-domain of recombinant human Factor VIII comprises 909 amino acids and is extensively N-and Oglycosylated, in that at least 20 different sites are occupied by numerous carbohydrate structures. This domain was incubated with trypsin and subjected to liquid chromatography electrospray ionization mass spectrometry analysis, using an electrospray orthogonal acceleration time-of-flight mass spectrometer as the detector for a capillary reversed phase HPLC separation of the digest. The inherent high mass resolution afforded by this instrument provides both ion charge state determination and high accuracy mass measurement that are of significant advantage in defining such highly complex mixtures.


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