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Received January 13, 2018
Accepted March 28, 2018
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Quantitative characterization of intact sialylated O-glycans with MALDI-MS for protein biotherapeutics
Cheol-Hwan Hwang
Hae-Min Park1
Han-Gyu Park
Da-Hee Ahn
Seong-Min Kim
Byoung Joon Ko2
Young Hwan Kim3
Yung-Hun Yang4
Yun-Gon Kim†
Department of Chemical Engineering, Soongsil University, Seoul 06978, Korea 1Department of Chemistry, Northwestern University, Evanston, Illinois 60208, U.S.A., USA 2New Drug Development Center, Osong Medical Innovation Foundation, Cheongju 28160, Korea 3Biomedical Omics Team, Korea Basic Science Institute (KBSI), Cheongju 28119, Korea 4Department of Biological Engineering, Konkuk University, Seoul 05029, Korea
Korean Journal of Chemical Engineering, July 2018, 35(7), 1462-1467(6), 10.1007/s11814-018-0058-0
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Abstract
For validating O-glycosylation of protein biotherapeutics, we presented a quantitative O-glycomics method which is based on the neutralization of sialic acids, the specific release of O-glycans, and the introduction of permanent positive charge followed by quantitative MALDI-MS analysis. This method shows excellent technical reproducibility, linearity and sensitivity. In addition, it enables the quantification of intact O-glycans with minimal degradation or loss of sialic acids on these glycans compared to a conventional HPLC-based method. We then applied this method to quantitatively characterize O-glycans present on Etanercept. The analysis showed the relative abundances of mono- and di-sialylated core 1 O-glycans - were 79.3±0.8% and 17.3±1.4%, respectively. This glycomics technology could allow for the reliable quantitative analysis of intact O-glycans from glycoproteins and may contribute to validation of O-glycosylation protein biotherapeutics in the pharmaceutical industry.
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