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 language | English (US) |
|---|---|
| Pages (from-to) | 7925-7935 |
| Number of pages | 11 |
| Journal | Analytical and Bioanalytical Chemistry |
| Volume | 406 |
| Issue number | 30 |
| DOIs | |
| State | Published - Nov 26 2014 |
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Keywords
- Human milk oligosaccharides
- Nano-flow liquid chromatography-mass spectrometry
- Rapid-throughput glycomics
ASJC Scopus subject areas
- Analytical Chemistry
- Biochemistry
Cite this
Rapid-throughput glycomics applied to human milk oligosaccharide profiling for large human studies. / Totten, Sarah M.; Wu, Lauren D.; Parker, Evan A.; Davis, Jasmine C C; Hua, Serenus; Stroble, Carol; Ruhaak, L. Renee; Smilowitz, Jennifer T.; German, J. Bruce; Lebrilla, Carlito B.
In: Analytical and Bioanalytical Chemistry, Vol. 406, No. 30, 26.11.2014, p. 7925-7935.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Rapid-throughput glycomics applied to human milk oligosaccharide profiling for large human studies
AU - Totten, Sarah M.
AU - Wu, Lauren D.
AU - Parker, Evan A.
AU - Davis, Jasmine C C
AU - Hua, Serenus
AU - Stroble, Carol
AU - Ruhaak, L. Renee
AU - Smilowitz, Jennifer T.
AU - German, J. Bruce
AU - Lebrilla, Carlito B
PY - 2014/11/26
Y1 - 2014/11/26
N2 - 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.]
AB - 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.]
KW - Human milk oligosaccharides
KW - Nano-flow liquid chromatography-mass spectrometry
KW - Rapid-throughput glycomics
UR - http://www.scopus.com/inward/record.url?scp=84913582609&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84913582609&partnerID=8YFLogxK
U2 - 10.1007/s00216-014-8261-2
DO - 10.1007/s00216-014-8261-2
M3 - Article
C2 - 25358913
AN - SCOPUS:84913582609
VL - 406
SP - 7925
EP - 7935
JO - Fresenius Zeitschrift fur Analytische Chemie
JF - Fresenius Zeitschrift fur Analytische Chemie
SN - 0016-1152
IS - 30
ER -