Quantitative characterization of intact sialylated O-glycans with MALDI-MS for protein biotherapeutics
| Autor: | Hae-Min Park, Yung-Hun Yang, Young Hwan Kim, Han-Gyu Park, Da-Hee Ahn, Cheol-Hwan Hwang, Seong-Min Kim, Yun-Gon Kim, Byoung Joon Ko |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
chemistry.chemical_classification Glycan Chromatography Maldi ms biology Chemistry General Chemical Engineering General Chemistry High-performance liquid chromatography Neutralization carbohydrates (lipids) Glycomics 03 medical and health sciences 030104 developmental biology O glycans biology.protein Glycoprotein Quantitative analysis (chemistry) |
| Zdroj: | Korean Journal of Chemical Engineering. 35:1462-1467 |
| ISSN: | 1975-7220 0256-1115 |
| DOI: | 10.1007/s11814-018-0058-0 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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