On-line CE-LIF-MS analysis of glycans labeled with teal fluorescence dye using electrokinetic sheath liquid pump-based nanospray ion source
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Authors
Khan, S.
Szabo, Z.
Liu, J.
Kunnummal, B.
Han, X.
Ouyang, Y.
Linhardt, Robert J.
Xia, Q.
Issue Date
2018
Type
Article
Language
ENG
Keywords
Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
Alternative Title
Abstract
Rationale: N-linked glycan analysis of recombinant therapeutic proteins, such as monoclonal antibodies, Fc-fusion proteins, and antibody-drug conjugates, provides valuable information regarding protein therapeutics glycosylation profile. Both qualitative identification and quantitative analysis of N-linked glycans on recombinant therapeutic proteins are critical analytical tasks in the biopharma industry during the development of a biotherapeutic. Methods: Currently, such analyses are mainly carried out using capillary electrophoresis/laser-induced fluorescence (CE/LIF), liquid chromatography/fluorescence (LC/FLR), and liquid chromatography/fluorescence/mass spectrometry (LC/FLR/MS) technologies. N-linked glycans are first released from glycoproteins by enzymatic digestion, then labeled with fluorescence dyes for subsequent CE or LC separation, and LIF or MS detection. Here we present an on-line CE/LIF/MS N-glycan analysis workflow that incorporates the fluorescent Teal™ dye and an electrokinetic pump-based anospray sheath liquid capillary electrophoresis/mass spectrometry (CE/MS) ion source. Results: Electrophoresis running buffer systems using ammonium acetate and ammonium hydroxide were developed for the negative ion mode CE/MS analysis of fluorescence-labeled N-linked glycans. Results show that on-line CE/LIF/MS analysis can be readily achieved using this versatile CE/MS ion source on common CE/MS instrument platforms. Conclusions: This on-line CE/LIF/MS method using Teal™ fluorescent dye and electrokinetic pump-based nanospray sheath liquid CE/MS coupling technology holds promise for on-line quantitation and identification of N-linked glycans on recombinant therapeutic proteins.
Description
Rapid Communications in Mass Spectrometry, 32, 882–888
Note : if this item contains full text it may be a preprint, author manuscript, or a Gold OA copy that permits redistribution with a license such as CC BY. The final version is available through the publisher’s platform.
Note : if this item contains full text it may be a preprint, author manuscript, or a Gold OA copy that permits redistribution with a license such as CC BY. The final version is available through the publisher’s platform.
Full Citation
On-line CE-LIF-MS analysis of glycans labeled with teal fluorescence dye using electrokinetic sheath liquid pump-based nanospray ion source, S. Khan, Z. Szabo, J. Liu, B. Kunnummal, X. Han, Y. Ouyang, R. J. Linhardt, Q. Xia, Rapid Communications in Mass Spectrometry, 32, 882–888, 2018.