Carbohydrate release from picomole quantities of glycoprotein and characterisation of glycans by high-performance liquid chromatography and mass spectrometry |
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| Institution: | 1. Department of Chemistry, University of Massachusetts Lowell, 1 University Avenue, Lowell, MA 01854, USA;2. Department of Chemistry, KTH-Royal Institute of Technology, Teknikringen 30, S-10044 Stockholm, Sweden;1. Key Laboratory of Marine Drugs, Ministry of Education, School of Medicine and Pharmacy, Ocean University of China, Shandong Provincial Key laboratory of Glycoscience and Glycoengineering, Qingdao 266003, China;2. Laboratory for Marine Drugs and Bioproducts of Qingdao National Laboratory for Marine Science and Technology, Qingdao 266237, China;3. Shandong Long Live Bio-Technology Co.,Ltd. Yucheng 251200, China;4. Shandong Key Laboratory of Straw and Stover Biorefinement Technologies, High-technology Development Zone, Yucheng, Shandong 251200, China;5. Shandong Functional Sugar Engineering Research Center, Yucheng, Shandong 251200, China;1. Key Laboratory of Marine Drugs, Shandong Provincial Key Laboratory of Glycoscience and Glycoengineering, School of Medicine and Pharmacy, Ocean University of China, Qingdao, Shandong, 266003, China;2. School of Industrial Technology, Universiti Sains Malaysia, Penang, 11800, Malaysia;1. Key Laboratory of Marine Drugs, Ministry of Education of China, School of Medicine and Pharmacy, Ocean University of China, Qingdao 266003, China;2. Shandong Provincial Key Laboratory of Glycoscience & Glycotechnology, Ocean University of China, Qingdao 266003, China;1. Faculty of Pharmaceutical Sciences, Josai University, 1-1 Keyakidai, Sakado, Saitama 350-0295, Japan;2. Technology Joint Management Office, Tokai University, 4-1-1 Kitakaname, Hiratsuka, Kanagawa 259-1292, Japan;3. Department of Human Ecology, Wayo Women’s University, Chiba 272-8533, Japan;4. Department of Applied Chemistry, School of Engineering, and Institute of Glycoscience, Tokai University, Kanagawa 259-1292, Japan;5. R&D Lab, NanoDex Inc., 2-3227 Mita, Kawasaki, Kanagawa 214-0034, Japan |
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| Abstract: | Samples of 5 to 20 μg of human IgG were subjected to dithiothreitol treatment to reduce disulphide bridges, followed by tryptic digestion. Glycans released from the tryptic peptide mixture by PNGase F digestion were then derivatised with 2-aminoacridone. Labelled oligosaccharides were separated by normal-phase high-performance liquid chromatography and individual components were collected for matrix-assisted laser desorption ionization time-of-flight and electrospray mass spectrometric analysis. |
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