SpringerProtocols: Abstract: Analysis of Neutral N-Linked Oligosaccharides From Antibodies Using Free-Solution Capillary Electrophoresis in Bare Fused-Silica Capillaries

Analysis of Neutral N-Linked Oligosaccharides From Antibodies Using Free-Solution Capillary Electrophoresis in Bare Fused-Silica Capillaries

By: Jeffrey S. Patrick³, Brenda P. Rener³, Gregory S. Clanton³, Avinash L. Lagu³

Abstract

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Conditions for the enzymatic release, chemical derivatization, and analysis of oligosaccharides from the consensus glycosylation sites on antibodies are described. Release of the oligosaccharides is from the native protein. The APTS derivatives of the released oligosaccharides are then analyzed by capillary electrophoresis (CE) using a free solution separation in a bare fused silica capillary. An example of the application of the method to the analysis of the oligosaccharide population from antibodies obtained from different cell lines is provided. The separation conditions provide for resolution of various galactose positional isomers, including those derived from different linkage configurations.

Affiliation(s): (3) Lilly Research Laboratories, Eli Lilly and Company, Lilly Corporate Center, Indianapolis, IN

Book Title: Capillary Electrophoresis of Proteins and Peptides

Series: Methods in Molecular Biology | Volume: 276 | Pub. Date: Jun-03-2004 | Page Range: 137-151 | DOI: 10.1385/1-59259-798-X:137

Subject: Biochemistry

Key Words: Antibodies - capillary zone electrophoresis - derivatization, fluorescence, fused-silica - LIF - N-linked - oligosaccharides - qualitative - quantitation - antibodies

References

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