A platform for high-throughput molecular characterization of recombinant monoclonal antibodies

Bailey, Mark J., Hooker, Andrew D., Adams, Carolyn S., Zhang, Shuhong and James, David C. (2005) A platform for high-throughput molecular characterization of recombinant monoclonal antibodies. Journal of Chromatography B, 826 1-2: 177-187. doi:10.1016/j.jchromb.2005.08.021


Author Bailey, Mark J.
Hooker, Andrew D.
Adams, Carolyn S.
Zhang, Shuhong
James, David C.
Title A platform for high-throughput molecular characterization of recombinant monoclonal antibodies
Journal name Journal of Chromatography B   Check publisher's open access policy
ISSN 1570-0232
Publication date 2005-11-01
Sub-type Article (original research)
DOI 10.1016/j.jchromb.2005.08.021
Volume 826
Issue 1-2
Start page 177
End page 187
Total pages 10
Publisher Elsevier B.V.
Language eng
Subject 03 Chemical Sciences
0301 Analytical Chemistry
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.
Keyword IgG2
Recombinant monoclonal antibody
Glycosylation
Glycopeptide
High-throughput characterization
Normal phase liquid chromatography
MALDI-TOF MS
Q-Index Code C1

Document type: Journal Article
Sub-type: Article (original research)
Collections: Excellence in Research Australia (ERA) - Collection
School of Chemical Engineering Publications
 
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Created: Wed, 19 Apr 2006, 20:39:00 EST