# A bioanalytical research program to unravel the human milk glycome

> **NIH NIH R35** · UTAH STATE HIGHER EDUCATION SYSTEM--UNIVERSITY OF UTAH · 2022 · $370,399

## Abstract

PROJECT SUMMARY
While human milk oligosaccharides (HMOs) are involved in a variety of biological processes that enable the
healthy development of the brain-gut axis of infants, a true understanding of how their molecular structures and
composition dictate such functions has largely remained a mystery. HMOs possess a tremendous degree of both
structural complexity and isomeric heterogeneity due to their many possible permutations of linkage positioning
and monosaccharide constituent arrangements. Therefore, improved analytical advancements are necessary to
unravel this isomerism puzzle and gain a better understanding of their infant-specific roles. Mass spectrometry
(MS) in conjunction with liquid chromatographic (LC) separations has been broadly used for HMO analyses.
Unfortunately, in many instances, LC-MS-based techniques suffer from insufficient chromatographic resolution
resulting in convoluted MS/MS spectra for co-eluting species, and thus limiting the accurate identification and
structural elucidation of HMOs. The overall goal of our research program is to develop a bioanalytical
workflow to enable more effective characterization of HMOs. To achieve this, we will implement a
multidimensional separations platform using recycling liquid chromatography coupled to high-
resolution cyclic ion mobility separations and mass spectrometry in conjunction with the development
of solution and gas-phase chemical probes and top-down sequencing approaches. Our multidimensional
separations platform will not only enable improved peak capacity, but also higher resolution separations to
facilitate the characterization of previously indistinguishable HMO isomers. Additionally, our cyclic ion mobility-
based separations will allow the determination of highly precise collision cross section values that will be
disseminated to other laboratories in the glycobiology community. Our chemical probes will facilitate the
improved identification of glycosidic linkage positioning in isomeric HMOs, especially when no authentic standard
is present. Lastly, our top-down sequencing strategies will enable the arrangement of monosaccharide
constituents to be better determined. Overall, we envision that our proposed bioanalytical toolbox will make
strides toward the de novo sequencing of HMOs and help achieve our overarching goal of unraveling the human
milk glycome to identify essential HMOs to be incorporated into infant formula.

## Key facts

- **NIH application ID:** 10497096
- **Project number:** 1R35GM146671-01
- **Recipient organization:** UTAH STATE HIGHER EDUCATION SYSTEM--UNIVERSITY OF UTAH
- **Principal Investigator:** Gabe Nagy
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $370,399
- **Award type:** 1
- **Project period:** 2022-09-21 → 2027-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10497096

## Citation

> US National Institutes of Health, RePORTER application 10497096, A bioanalytical research program to unravel the human milk glycome (1R35GM146671-01). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10497096. Licensed CC0.

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