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Coupling porous sheathless interface MS with transient‐ ITP in neutral capillaries for improved sensitivity in glycopeptide analysis
Author(s) -
Heemskerk Anthonius A. M.,
Wuhrer Manfred,
Busnel JeanMarc,
Koeleman Carolien A. M.,
Selman Maurice H. J.,
Vidarsson Gestur,
Kapur Rick,
Schoenmaker Bart,
Derks Rico J. E.,
Deelder André M.,
Mayboroda Oleg A.
Publication year - 2013
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201200357
Subject(s) - chemistry , glycosylation , glycopeptide , glycoprotein , chromatography , glycan , biophysics , biochemistry , antibiotics , biology
I g G antibodies are modulated in their function by the specific structure of the N ‐glycans attached to their F c (fragment crystallizable) portions. However, the glycosylation analysis of antigen‐specific I g G s is a challenging task as antibody levels to a given antigen only represent a fraction of the total I g G levels. Here, we investigated the use of a transient‐ ITP (t‐ ITP )— MS method for highly sensitive I g G 1 glycosylation profiling as a complementary method to a high‐throughput nano‐ RPLC ‐ MS method. It was found that t‐ ITP ‐ CZE using neutrally coated separation capillaries with a large volume injection (37% of capillary volume) and interfaced to MS with a sheathless porous sprayer yielded a 40‐fold increase in sensitivity for I g G 1 F c glycopeptide analysis when compared to the conventional strategy. Furthermore, the glycoform profiles found with the t‐ ITP ‐ CZE strategy were comparable to those from nano‐ RPLC ‐ MS . In conclusion, the use of the highly sensitive t‐ ITP ‐ CZE ‐ MS method will provide information on I g G F c glycosylation for those samples with IgG1 concentrations below the LOD s of the conventional method.