# Engineered probes for sialoglycan detection

> **NIH NIH R01** · VANDERBILT UNIVERSITY · 2022 · $451,153

## Abstract

PROJECT SUMMARY/ABSTRACT
 Sialoglycans can be poor antigens, meaning that it can be challenging to develop effective antibodies for
their detection. Instead, sialoglycan mapping heavily relies upon mass spectrometry, which requires expertise
and instrumentation available to few laboratories. One recent push in the field has been to harvest the breadth
of selectivity found within naturally-occurring sialoglycan-binding proteins in order to develop glycan-detecting
proteins. This strategy identified a number of sialoglycan-binding proteins, particularly those that bind with high
affinity and narrow selectivity to sialyl-T antigen (sTa). However, major gaps remain in the spectrum of the
sialoglycans that are recognized.
 Here, we focus on sialoglycans that are likely abundant on cells or that are prevalent in disease but that lack
practical probes for their detection, specifically α2,3 linked and α2,6 linked sialoglycans. We propose to create
probes that recognize these glycans by tailoring the specificity of existing sialoglycan binding proteins using
structure-based protein engineering.
 In Aim 1, we engineer the bacterial Siglec-like adhesins to create a library of probes for tri- and tetra-
saccharides, each of which binds one α2,3 linked sialoglycan with high affinity and narrow selectivity.
 In Aim 2, we apply engineering principles to the development of probes for α2,3 linked sialoglycans and
focus on the α2,6 linked sialyl Tn antigen disaccharide, a biomarker for cancer.
 In Aim 3, we evaluate the utility of these probes in measuring glycans in human plasma, and cross-validated
these by affinity capture and mass spectrometry. The ability to distinguish glycan abundance and repertoire in
healthy donors versus cancer patients will be included as a part of this aim. Successful probes will be distributed
for use both in lectin arrays and in low-throughput assays.

## Key facts

- **NIH application ID:** 10438835
- **Project number:** 5R01GM137458-03
- **Recipient organization:** VANDERBILT UNIVERSITY
- **Principal Investigator:** T M Iverson
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $451,153
- **Award type:** 5
- **Project period:** 2020-09-20 → 2024-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10438835

## Citation

> US National Institutes of Health, RePORTER application 10438835, Engineered probes for sialoglycan detection (5R01GM137458-03). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10438835. Licensed CC0.

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