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|
| Type |
Poster Presentation |
| Area |
Analytical Chemistry |
| Room No. |
Exhibition Hall 2+3 |
| Time |
10월 19일 (목요일) 11:00~12:30 |
| Code |
ANAL.P-167 |
| Subject |
High-throughput Automated Platform for Native Glycan Analysis using Liquid Handling System |
| Authors |
Gyeong Mi Park, Hyun Joo An*
Graduate School of Analytical Science and Technology, Chungnam National University, Korea
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| Abstract |
Glycosylation of therapeutic glycoproteins is a key factor influencing biological function, safety, and protein stability. To assure quality of biopharmaceuticals, glycosylation should be analyzed at all stages of manufacturing process since it could be changed by cell culture and environmental conditions. For sample preparation, automated system is often necessary in pharmaceutical industry to reduce hands-on time and to obtain accurate and reproducible data. Here, we present the robotic platform to release and enrich glycans from glycoproteins by liquid handling system combined with 96-well microplate. Overall processes including protein denaturation by heating block, N-glycan release by PNGase F, and glycan enrichment by solid phase extraction (SPE) were automatically performed. Especially, the technical variation was mostly affected by PGC-SPE to purify and fractionate different types of glycans. For setting up SPE conditions, we used glycoprotein mixtures including diverse glycans which were eluted by varying proportions of acetonitrile in water and formic acid. Consequently, we obtained three eluents consisting of high-mannose and complex-type glycans w/wo sialic acid residues. Automated full steps for glycan preparation is being repeatedly performed to validate the technical reproducibility of our platform. Ultimately, it can be applied for monitoring bioactive glycans of biotherapeutics or clinical glycan biomarkers. |
| E-mail |
pkm2432@gmail.com |
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120th General Meeting of the KCS