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Rapid-throughput glycomics applied to human milk oligosaccharide profiling for large human studies

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Abstract

Glycomic analysis is the comprehensive determination of glycan (oligosaccharide) structures with quantitative information in a biological sample. Rapid-throughput glycomics is complicated due to the lack of a template, which has greatly facilitated analysis in the field of proteomics. Furthermore, the large similarities in structures make fragmentation spectra (as obtained in electron impact ionization and tandem mass spectrometry) less definitive for identification as it has been in metabolomics. In this study, we develop a concept of rapid-throughput glycomics on human milk oligosaccharides, which have proven to be an important bioactive component of breast milk, providing the infant with protection against pathogenic infection and supporting the establishment of a healthy microbiota. To better understand the relationship between diverse oligosaccharides structures and their biological function as anti-pathogenic and prebiotic compounds, large human studies are needed, which necessitate rapid- to high-throughput analytical platforms. Herein, a complete glycomics methodology is presented, evaluating the most effective human milk oligosaccharide (HMO) extraction protocols, the linearity and reproducibility of the nano-liquid chromatography chip time-of-flight mass spectrometry (nano-LC chip-TOF MS) method, and the efficacy of newly developed, in-house software for chromatographic peak alignment that allows for rapid data analysis. High instrument stability and retention time reproducibility, together with the successful automated alignment of hundreds of features in hundreds of milk samples, allow for the use of an HMO library for rapid assignment of fully annotated structures.

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Acknowledgments

The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Research reported in this publication was supported by the National Institute of Child Health and Human Development, National Institute of General Medicine, and National Center of Complementary and Alternative Medicine of the National Institutes of Health under award numbers R01HD061923, R01GM049077, R01 AT007079, and 1U24DK097154.

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Authors and Affiliations

  1. Department of Chemistry, University of California, Davis, CA, 95616, USA

    Sarah M. Totten, Lauren D. Wu, Evan A. Parker, Jasmine C. C. Davis, Serenus Hua, Carol Stroble, L. Renee Ruhaak & Carlito B. Lebrilla

  2. Asia Glycomics Reference Site, Chungnam National University, Daejeon, 305-764, South Korea

    Serenus Hua

  3. Foods for Health Institute, University of California, Davis, CA, 95616, USA

    Jennifer T. Smilowitz & J. Bruce German

  4. Department of Food Science & Technology, University of California, Davis, CA, 95616, USA

    Jennifer T. Smilowitz & J. Bruce German

Authors
  1. Sarah M. Totten
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  2. Lauren D. Wu
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  3. Evan A. Parker
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  4. Jasmine C. C. Davis
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  5. Serenus Hua
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  6. Carol Stroble
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  7. L. Renee Ruhaak
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  8. Jennifer T. Smilowitz
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  9. J. Bruce German
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  10. Carlito B. Lebrilla
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Corresponding author

Correspondence to Carlito B. Lebrilla.

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Totten, S.M., Wu, L.D., Parker, E.A. et al. Rapid-throughput glycomics applied to human milk oligosaccharide profiling for large human studies. Anal Bioanal Chem 406, 7925–7935 (2014). https://doi.org/10.1007/s00216-014-8261-2

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  • DOI: https://doi.org/10.1007/s00216-014-8261-2

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