Highly sensitive glycosylation analysis of membrane glycoproteins avoiding polymeric contaminants |
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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 |
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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
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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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