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Highly sensitive glycosylation analysis of membrane glycoproteins avoiding polymeric contaminants

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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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Authors and Affiliations

  1. School of Chemical and Biological Engineering, Seoul National University, Seoul, 151-742, Korea

    Hee-Jin Jeong & Hae-Min Park

  2. Institute of Bioengineering, Seoul National University, Seoul, 151-742, Korea

    Byung-Gee Kim

  3. Department of Chemical Engineering, Soongsil University, Seoul, 156-743, Korea

    Kyoung-Jin Kim, Yoon-Woo Kim & Yun-Gon Kim

  4. Department of Microbial Engineering, College of Engineering, Konkuk University, Seoul, 143-701, Korea

    Shashi Kant Bhatia & Yung-Hun Yang

  5. Arctic Research Center, Korea Polar Research Institute, KIOST, Incheon, 406-840, Korea

    Yoo Kyung Lee

Authors
  1. Hee-Jin Jeong
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  2. Hae-Min Park
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  3. Kyoung-Jin Kim
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  4. Yoon-Woo Kim
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  5. Shashi Kant Bhatia
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  7. Yung-Hun Yang
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  8. Byung-Gee Kim
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  9. Yun-Gon Kim
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Correspondence to Yun-Gon Kim.

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Jeong, HJ., Park, HM., Kim, KJ. et al. Highly sensitive glycosylation analysis of membrane glycoproteins avoiding polymeric contaminants. Biotechnol Bioproc E 19, 545–550 (2014). https://doi.org/10.1007/s12257-014-0117-2

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  • DOI: https://doi.org/10.1007/s12257-014-0117-2

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