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

Sarah M. Totten, Lauren D. Wu, Evan A. Parker, Jasmine C C Davis, Serenus Hua, Carol Stroble, L. Renee Ruhaak, Jennifer T. Smilowitz, J. Bruce German, Carlito B Lebrilla

Research output: Contribution to journalArticle

30 Scopus citations

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. [Figure not available: see fulltext.]

Original languageEnglish (US)
Pages (from-to)7925-7935
Number of pages11
JournalAnalytical and Bioanalytical Chemistry
Volume406
Issue number30
DOIs
StatePublished - Nov 26 2014

Keywords

  • Human milk oligosaccharides
  • Nano-flow liquid chromatography-mass spectrometry
  • Rapid-throughput glycomics

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry

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    Totten, S. M., Wu, L. D., Parker, E. A., Davis, J. C. C., Hua, S., Stroble, C., Ruhaak, L. R., Smilowitz, J. T., German, J. B., & Lebrilla, C. B. (2014). Rapid-throughput glycomics applied to human milk oligosaccharide profiling for large human studies. Analytical and Bioanalytical Chemistry, 406(30), 7925-7935. https://doi.org/10.1007/s00216-014-8261-2