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| Type |
Poster Presentation |
| Area |
Analytical Chemistry |
| Room No. |
Event Hall |
| Time |
4월 19일 (목요일) 11:00~12:30 |
| Code |
ANAL.P-230 |
| Subject |
A Simultaneous Determination of Highly Acidic Glycans in Biotherapeutics using PGC-SPE and LC-MS/MS
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| Authors |
Youngsuk Seo, Hyun Joo An*
Graduate School of Analytical Science and Technology, Chungnam National University, Korea
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| Abstract |
Glycosylation of therapeutic glycoproteins influences on pharmacological functions including efficacy, safety, and biological activity. In a therapeutic enzyme, phosphorylated glycans play a significant role as the target signal to enter into the lysosome. Sialylated glycans are also an important bioactive glycan which can improve in vivo recovery of a therapeutic enzyme. Therefore, the in-depth analysis of bioactive glycans has been required to ensure product consistency with the increase of the biosimilars market. However, glycosylation characterization of a therapeutic enzyme is still challengeable because of an increased heterogeneity by phosphorylated glycosylation as a modified glycoform and an extended sialylation. Here, we present an analytical approach to readily determine phosphorylated glycans and sialylated glycans, individually using porous graphitized carbon (PGC)-solid phase extraction (SPE). We successfully fractionated them according to varying proportions of acetonitrile in water and acid, though both phosphorylated glycans and sialylated glycans showed a high acidity. Each eluent was analyzed by LC/MS and CID MS/MS to fully characterize glycan composition, detailed structure, and relative quantitation. We observed mannose-phosphate series consisting of Man 5 to Man 9 decorated with mono-or di-phosphate groups. Sialylation of a therapeutic enzyme was mainly composed of glycans having two NeuAc residues. Bioactive glycan analysis of an original therapeutic enzyme and biosimilar is being further performed to demonstrate our analytical platform as an assessment tool for glycosylation biosimilarity.
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| E-mail |
ysseo910@gmail.com |
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121st General Meeting of the KCS