# Mass Spectrometry-Based Biochemical Analysis of Single Cells Beyond the Global Proteome

> **NIH NIH R35** · BRIGHAM YOUNG UNIVERSITY · 2024 · $469,543

## Abstract

PROJECT SUMMARY/ABSTRACT
Biological tissues exhibit a high degree of phenotypic heterogeneity and plasticity, comprising many different
subpopulations of cells in various states. Quantifying this heterogeneity at the single-cell level and with molecular
depth across large numbers of cells and multiple classes of molecules provides information that cannot be
obtained at the bulk scale and will ultimately lead to improved diagnostics and more effective treatments. While
single-cell nucleic acid sequencing approaches are having a significant impact on biomedical research, proteins,
lipids and metabolites mediate the bulk of cellular function and measurement of their expression provides more
direct insight into cellular phenotype. There is thus an urgent need to develop new technologies for large-scale
direct proteome, lipidome and metabolome profiling at the single-cell level. To fill this gap, mass spectrometry
(MS)-based profiling of protein expression in single cells has recently been demonstrated through the
implementation of more efficient sample processing workflows, novel experimental designs and improved
instrument sensitivity. Label-free MS-based proteomics can now quantify >3,000 protein groups per cell across
>4 orders of magnitude of dynamic range. Here we propose to apply mass spectrometry to study biomolecular
expression at the single-cell level beyond the global proteome. We will develop global and targeted approaches
to profile posttranslational modifications in single cells, beginning with phosphorylation. We will also extend
nanoflow liquid chromatography-MS capabilities for in-depth single-cell lipid profiling. Ultimately, we will develop
novel means of generating complex LC gradients that utilize more than two mobile phases to efficiently profile
multiple classes of biomolecules (e.g., proteome and lipidome) from the same single cell. These research
directions will, in combination with mature nucleic acid sequencing strategies, provide an unprecedented view of
cellular regulation from genotype to phenotype at the single-cell level.

## Key facts

- **NIH application ID:** 10836936
- **Project number:** 1R35GM153179-01
- **Recipient organization:** BRIGHAM YOUNG UNIVERSITY
- **Principal Investigator:** Ryan T Kelly
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $469,543
- **Award type:** 1
- **Project period:** 2024-09-01 → 2029-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10836936

## Citation

> US National Institutes of Health, RePORTER application 10836936, Mass Spectrometry-Based Biochemical Analysis of Single Cells Beyond the Global Proteome (1R35GM153179-01). Retrieved via AI Analytics 2026-05-28 from https://api.ai-analytics.org/grant/nih/10836936. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*