# High performance and widely accessible secretomics assay platform

> **NIH NIH R44** · AURAGENT BIOSCIENCE, LLC · 2022 · $943,531

## Abstract

Abstract
 Secretory proteins are fundamental to local and systemic cellular signaling in biological systems.
The regulation and dysregulation of the networks involving these proteins plays a critical role in
health, aging, and development of disease. As such, study of these proteins and their networks
is critical to our understanding of biology. Not surprisingly, this particular set of proteins constitutes
a large proportion of the targets of pharmaceutical drugs, and also represents attractive targets
for diagnostics and future precision medicine efforts. Unfortunately, many of these secreted
proteins are difficult to detect. There are existing technologies for studying these secreted proteins
but they either underperform, are extremely expensive and/or complex given their specific-utility.
containing multiple cell types. Although the need to study secreted proteins is ubiquitous, there
is not a technology that is widely available (inexpensive) and capable of providing
information on multiple secreted proteins simultaneously at single-cell resolution, and we
aim to address this need in the current project.
 At the core of our technologies is the Plasmonic Fluor (PF), our proprietary, ultrabright
fluorescent nanoconstruct designed to enhance the performance of fluorescence-based
immunoassays. We have already built a high-sensitivity, multiplexed immunoassay platform
leveraging this technology, which is currently being tested with external collaborators. This
platform includes an inexpensive, high performance, single-laser fluorescence microplate/slide
reader optimized for use with our PFs. We will extend this existing platform to accommodate PF-
enhanced cellular secretion assays we call PFluorospot. The upgraded platform that will result
from the proposed effort will be a truly versatile and powerful tool for the measurement of
proteins.
 In our preliminary work, we have successfully manufactured at commercial scale and with high
reproducibility streptavidin-conjugated PFs that are spectrally equivalent to FITC, Cy3, and Cy5
(PF490, PF550, and PF650, respectively). We have also created three PFs with entirely unique
excitation/emission spectra: two which have excitation spectra like FITC and emit like Cy3 and
Cy5 (PF490_550 and PF490_650, respectively), and one which has an excitation spectrum like Cy3 but
emits like Cy5 (PF550_650). These unique PFs were synthesized at a prototype scale which we aim
to scale up in this effort because they will allow 3 lasers to be used to interrogate 6 separate
targets. In a demonstration of applicability to cellular secretion assays, streptavidin-PF550 and
PF650 were conjugated to antibodies at a small scale and used to perform dual-color PFluorospot.
These initial PFluorospot experiments show that additional information can be obtained in these
assays relative to ELISPOT/FluoroSpot, and are simpler and significantly faster to perform. It was
possible to visualize individual cells for adherent cells and better...

## Key facts

- **NIH application ID:** 10547532
- **Project number:** 1R44GM148138-01
- **Recipient organization:** AURAGENT BIOSCIENCE, LLC
- **Principal Investigator:** Scott L Crick
- **Activity code:** R44 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $943,531
- **Award type:** 1
- **Project period:** 2022-09-17 → 2024-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10547532

## Citation

> US National Institutes of Health, RePORTER application 10547532, High performance and widely accessible secretomics assay platform (1R44GM148138-01). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10547532. Licensed CC0.

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