Characterization of glyco isoforms in plasma-derived human antithrombin by on-line capillary zone electrophoresis-electrospray ionization-quadrupole ion trap-mass spectrometry of the intact glycoproteins

Electrophoresis
Uwe DemelbauerAndreas Rizzi

Abstract

The carbohydrate structures of five isoforms of alpha-AT and two isoforms of beta-AT were determined by applying capillary zone electrophoresis (CZE) on-line coupled to electrospray ionization-mass spectrometry (ESI-MS) using an ion-trap analyzer. For the AT preparations gained from a plasma pool at least semiquantitative information on the isoform-distributions could be gained. Unlike to the commonly used approaches starting from enzymatically treated glycoproteins, this approach deals with intact proteins. The high accuracy of the molecular mass determination obtained by the ion-trap analyzer allows one to calculate and ascertain the carbohydrate composition assuming no variations in the protein moiety of AT and to exclude or confirm the presence of the potential post-translational or other modifications. Therefore, the direct coupling of CZE with ESI-MS does not only represent a fast alternative technique to two-dimensional electrophoresis (2-DE) but serves as a method which provides structural information complementary to that gained from peptide mapping methods.

Citations

Jul 10, 2010·Analytical and Bioanalytical Chemistry·Sara Ongay, Christian Neusüss
Jul 15, 2005·Journal of Chromatography. a·Uwe M DemelbauerAndreas Rizzi
Jan 29, 2005·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Maria Rowena N Monton, Shigeru Terabe
Jan 27, 2005·Journal of Mass Spectrometry : JMS
Jul 6, 2011·Journal of Chromatography. a·Ruchi GuptaBruce D Grieve
Jun 15, 2007·Journal of Chromatography. a·Rob HaselbergGovert W Somsen
Oct 20, 2006·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Manfred WuhrerCornelis H Hokke
Nov 14, 2006·Journal of Chromatography. a·Javier Hernández-BorgesAlejandro Cifuentes
Mar 19, 2005·Electrophoresis·Philippe Schmitt-Kopplin, Matthias Englmann
Jan 20, 2006·Electrophoresis·Suresh Babu C VYoung Sook Yoo
Nov 25, 2005·Electrophoresis·Vladislav Dolník
Mar 13, 2008·Mass Spectrometry Reviews·Caterina TemporiniGabriele Caccialanza
May 25, 2007·Electrophoresis·Victoria Sanz-NebotJosé Barbosa
Nov 12, 2015·Glycoconjugate Journal·Florent ClercManfred Wuhrer
Dec 30, 2017·Expert Review of Proteomics·Arun V Everest-DassNicolle H Packer
Aug 20, 2019·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Yuki YagiKatsuyoshi Yamazaki
Nov 2, 2019·Journal of Pharmaceutical and Biomedical Analysis·Julien CamperiNathalie Delaunay
Jun 15, 2006·Analytical Chemistry·James KralyNorman J Dovichi

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