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The effects of abundant plasma protein depletion on global glycan profiling using NanoLC FT-ICR mass spectrometry

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Abstract

We report the results of abundant plasma protein depletion on the analysis of underivatized N-linked glycans derived from plasma proteins by nanoLC Fourier-transform ion cyclotron resonance mass spectrometry. N-linked glycan profiles were compared between plasma samples where the six most abundant plasma proteins were depleted (n = 3) through a solid-phase immunoaffinity column and undepleted plasma samples (n = 3). Three exogenous glycan standards were spiked into all samples which allowed for normalization of the N-glycan abundances. The abundances of 20 glycans varying in type, structure, composition, and molecular weight (1,200–3,700 Da) were compared between the two sets of samples. Small fucosylated non-sialylated complex glycans were found to decrease in abundance in the depleted samples (greater than or equal to tenfold) relative to the undepleted samples. Protein depletion was found to marginally effect (less than threefold) the abundance of high mannose, hybrid, and large highly sialylated complex species. The significance of these findings in terms of future biomarker discovery experiments via global glycan profiling is discussed.

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Acknowledgments

The authors gratefully acknowledge the financial support provided by the National Institutes of Health (R33 CA105295), the W.M. Keck Foundation, and North Carolina State University.

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Authors and Affiliations

  1. W.M. Keck FT-ICR Mass Spectrometry Laboratory, Department of Chemistry, North Carolina State University, Raleigh, NC, 27695, USA

    Michael S. Bereman & David C. Muddiman

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  1. Michael S. Bereman
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  2. David C. Muddiman
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Correspondence to David C. Muddiman.

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Bereman, M.S., Muddiman, D.C. The effects of abundant plasma protein depletion on global glycan profiling using NanoLC FT-ICR mass spectrometry. Anal Bioanal Chem 396, 1473–1479 (2010). https://doi.org/10.1007/s00216-009-3368-6

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  • DOI: https://doi.org/10.1007/s00216-009-3368-6

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