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Evaluation of Desialylation during 2-Amino Benzamide Labeling of Asparagine-Linked Oligosaccharides

Authors
  • Aich, Udayananth
  • Hurum, Deanna C.
  • Basumallick, Lipika
  • Rao, Srinivasa
  • Pohl, Chris
  • Rohrer, Jeffrey S.
  • Kandzia, Sebastian
Type
Published Article
Journal
Analytical Biochemistry
Publication Date
Jan 01, 2014
Accepted Date
Mar 13, 2014
Identifiers
DOI: 10.1016/j.ab.2014.03.008
Source
Elsevier
Keywords
License
Unknown

Abstract

Labeling of released asparagine-linked (N-linked) oligosaccharides from glycoproteins is commonly performed to aid in the separation and detection of the oligosaccharide. Of the many available oligosaccharide labels, 2-amino benzamide (2-AB) is a popular choice for providing a fluorescent product. The derivatization conditions can potentially lead to oligosaccharide desialylation. This work evaluates the extent of sialic acid loss during 2-AB labeling of N-linked oligosaccharides released from bovine fetuin, polyclonal human serum IgG, and human α1-acid glycoprotein as well as of sialylated oligosaccharide reference standards and finds that for more highly sialylated oligosaccharides the loss is greater than the <2% value commonly cited. Manufacturers of glycoprotein biotherapeutics need to produce products with a consistent state of sialylation, and therefore require an accurate assessment of glycoprotein sialylation.

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