| Article ID | Journal | Published Year | Pages | File Type |
|---|---|---|---|---|
| 9753205 | Journal of Chromatography B | 2005 | 11 Pages |
Abstract
We describe quantitative characterization of a sample preparation platform for rapid and high-throughput analysis of recombinant monoclonal antibodies (MAbs) and their post-translational modifications. MAb capture, desalting and in situ reduction/alkylation were accomplished by sequential adsorption of analyte to solid phase beads (protein A, reverse-phase) suspended in microtiter plate wells. Following elution and rapid tryptic digestion in the presence of acid-labile surfactant (RapiGestâ¢), peptides were fractionated by stepwise elution from reverse-phase pipet tips and the fraction containing Fc N-glycopeptides isolated. Direct quantitative analysis of the relative abundance of peptide glycoforms by MALDI-TOF MS in linear mode closely correlated with normal phase HPLC analysis of fluorophore labeled N-glycans released by PNGaseF.
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Physical Sciences and Engineering
Chemistry
Analytical Chemistry
Authors
Mark J. Bailey, Andrew D. Hooker, Carolyn S. Adams, Shuhong Zhang, David C. James,