Rapid magnetic bead based sample preparation for automated and high throughput N-glycan analysis of therapeutic antibodies

Csaba Váradi, Clarence Lew, A. Guttman

Research output: Contribution to journalArticle

47 Citations (Scopus)

Abstract

Full automation to enable high throughput N-glycosylation profiling and sequencing with good reproducibility is vital to fulfill the contemporary needs of the biopharmaceutical industry and requirements of national regulatory agencies. The most prevalently used glycoanalytical methods of capillary electrophoresis and hydrophilic interaction liquid chromatography, while very efficient, both necessitate extensive sample preparation and cleanup, including glycoprotein capture, N-glycan release, fluorescent derivatization, purification, and preconcentration steps during the process. Currently used protocols to fulfill these tasks require multiple centrifugation and vacuum-centrifugation steps, making liquid handling robot mediated automated sample preparation difficult and expensive. In this paper we report on a rapid magnetic bead based sample preparation approach that enables full automation including all the process phases just in a couple of hours without requiring any centrifugation and/or vacuum centrifugation steps. This novel protocol has been compared to conventional glycan sample preparation strategies using standard glycoproteins (IgG, fetuin, and RNase B) and featured rapid processing time, high release and labeling efficiency, good reproducibility, and the potential of easy automation.

Original languageEnglish
Pages (from-to)5682-5687
Number of pages6
JournalAnalytical Chemistry
Volume86
Issue number12
DOIs
Publication statusPublished - Jun 17 2014

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Centrifugation
Polysaccharides
Automation
Throughput
Antibodies
Vacuum
Fetuin-B
Glycoproteins
Glycosylation
Capillary electrophoresis
Liquid chromatography
Capillary Electrophoresis
Therapeutics
Hydrophobic and Hydrophilic Interactions
Liquid Chromatography
Labeling
Purification
Industry
Immunoglobulin G
Robots

ASJC Scopus subject areas

  • Analytical Chemistry
  • Medicine(all)

Cite this

Rapid magnetic bead based sample preparation for automated and high throughput N-glycan analysis of therapeutic antibodies. / Váradi, Csaba; Lew, Clarence; Guttman, A.

In: Analytical Chemistry, Vol. 86, No. 12, 17.06.2014, p. 5682-5687.

Research output: Contribution to journalArticle

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