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Highly sensitive glycosylation analysis of membrane glycoproteins avoiding polymeric contaminants
Authors:Hee-Jin Jeong  Hae-Min Park  Kyoung-Jin Kim  Yoon-Woo Kim  Shashi Kant Bhatia  Yoo Kyung Lee  Yung-Hun Yang  Byung-Gee Kim  Yun-Gon Kim
Affiliation:1. School of Chemical and Biological Engineering, Seoul National University, Seoul, 151-742, Korea
3. Department of Chemical Engineering, Soongsil University, Seoul, 156-743, Korea
4. Department of Microbial Engineering, College of Engineering, Konkuk University, Seoul, 143-701, Korea
5. Arctic Research Center, Korea Polar Research Institute, KIOST, Incheon, 406-840, Korea
2. Institute of Bioengineering, Seoul National University, Seoul, 151-742, Korea
Abstract:We established a ‘seize-and-release’ purification method to eliminate polyhexose contaminants for a highly sensitive glycan profiling. Pig liver membrane lysates were pretreated with sodium dodecyl sulfate (SDS) surfactant and subsequently dialyzed to remove polyhexose contaminants. From the purified membrane glycoproteins, glycans were released and identified by mass spectrometry. As a result, we clearly obtained N- and O-glycan profiles of a pig liver, which were not achieved without any pre-treatments. This technique demonstrates a powerful approach for enhancing the sensitivity of MS-based glycan profiling.
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