# Multiplexed analysis of secreted proteins from single-cells using high dynamic range nanovials

> **NIH NIH R43** · PARTILLION BIOSCIENCE CORPORATION · 2023 · $275,489

## Abstract

ABSTRACT
Cellular secretions make up a significant portion of the proteome, playing a crucial role in governing a range of
bodily functions – in particular immunity. Despite their significance, the study at cellular secretions at the single-
cell level has been limited due to a lack of accessible technologies. An ideal assay would have single-cell
resolution, measure several secreted products, and allow for recovery of cells for further downstream study. No
one assay fulfills all of these requirements. The lack of methodologies to identify and interrogate cells which
display high levels of secretory capacity has effectively stalled research into these potentially highly potent cells
for cell therapies. The most common technique to characterize cell secretion at the single-cell level is the
decades old ELISpot technology, however, this technology is limited in the number of secretions that can be
measured and cells cannot be sorted based on the results of the assay. Thus, there is a critical need to develop
accessible technologies which offer the capability to functionally screen and recover cells in a highly multiplexed
secretion format. To address this gap, we will engineer nanovials, cavity-containing hydrogel particles, to create
a robust 5-plex assay format capable of measuring five secreted cytokines from single cells, supporting several
different workflows such as rare polyfunctional T cell sorting, or other immune cell screening by function. In the
first aim of this project, we will develop nanovial formulations compatible with a 5-plex secretion assay by
increasing the number of binding sites for secretion capture. We will also identify suitable antibody pairs and
modification stoichiometry for IFN-gamma, TNF-alpha, GM-CSF, IL-2, and Granzyme B. In the second aim we
will validate the nanovial single-cell assays by comparing the single-cell secretions of HLA-matched PBMCs
activated by peptide presentation using both ELISpot and the developed nanovial assays. Successful completion
of our aims will address critical gaps in traditional secretion-based profiling of single cells such as ELISpot or
intracellular cytokine staining. This multiplexed assay will enable researchers to discriminate and recover highly
functional cells which will spur innovation in both fundamental research and therapeutic development.

## Key facts

- **NIH application ID:** 10761557
- **Project number:** 1R43AI179363-01
- **Recipient organization:** PARTILLION BIOSCIENCE CORPORATION
- **Principal Investigator:** Joseph de Rutte
- **Activity code:** R43 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $275,489
- **Award type:** 1
- **Project period:** 2023-08-01 → 2024-01-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10761557

## Citation

> US National Institutes of Health, RePORTER application 10761557, Multiplexed analysis of secreted proteins from single-cells using high dynamic range nanovials (1R43AI179363-01). Retrieved via AI Analytics 2026-05-29 from https://api.ai-analytics.org/grant/nih/10761557. Licensed CC0.

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