Highly sensitive glycosylation analysis of membrane glycoproteins avoiding polymeric contaminants

• Published in: Biotechnology and bioprocess engineering Vol. 19; no. 3; pp. 545 - 550
• Main Authors: Jeong, Hee-Jin, Park, Hae-Min, Kim, Kyoung-Jin, Kim, Yoon-Woo, Bhatia, Shashi Kant, Lee, Yoo Kyung, Yang, Yung-Hun, Kim, Byung-Gee, Kim, Yun-Gon
• Format: Journal Article
• Language: English
• Published: Heidelberg Springer-Verlag 01.06.2014; The Korean Society for Biotechnology and Bioengineering; Springer Nature B.V; 한국생물공학회
• Subjects: Bioengineering; Biotechnology; Caustic soda; Chemistry; Chemistry and Materials Science; Contaminants; Engineering; Glycoproteins; Glycosylation; Hemodialysis; Hogs; Industrial and Production Engineering; Ions; Lasers; Lipids; Liver; Mass spectrometry; membrane glycoproteins; polysaccharides; Process engineering; purification methods; R&D; Research & development; Research Paper; Scientific imaging; Sodium; sodium dodecyl sulfate; Studies; Surfactants; swine; 생물공학; South Korea; MALDI-TOF MS; glycan; sodium dodecyl sulfate (SDS) surfactant; polyhexose contaminants
• ISSN: 1226-8372; 1976-3816
• DOI: 10.1007/s12257-014-0117-2

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.