# Administrative Equipment Supplement to Enabling Technology to Screen and Quantify Sialylated Structures for Activity Against Viral Enzymes and Receptors

> **NIH NIH R01** · WEST VIRGINIA UNIVERSITY · 2021 · $205,000

## Abstract

Abstract for R01GM140560: Sialylated glycans are involved in complex regulation and signaling, and
play a critical role in disease. In the virus life cycle receptor binding and sialic acid cleavage to facilitate
release can compete and are therefore delicately balanced. While monovalent binding with a single
sialylated ligand is weak (KD ~mM), hemagglutinin forms a trimer, which enables multivalent binding.
Multivalent binding involving more than 1 ligand leads to strong binding (KD ~nM). This switch between
multivalent and monovalent binding allows the hemagglutinin to bind to the host with high affinity that
is easily reversed following replication and release by cleavage of only some of the sialylated ligands.
Neuraminidase inhibitors for influenza (Tamiflu, Relenza, and Rapivab) block the neuraminidase
cleavage in some infections. The development of therapeutic strategies to block hemagglutinin binding
with small molecule sialylated inhibitors has been limited. Several analytical barrier s to the analysis of
sialic acids and sialylated compounds have challenged research in this area. The long term objective
of this project is to bridge this gap with enabling technology through two key innovations. A new
screening approach for enzymes and receptors is introduced through the use of thermally reversible
nanogels. With this new strategy the sialic acid structures that interact with enzymes or receptors are
identified through capillary electrophoresis. This work is based on rapid-in line exoglycosidase
reactions facilitated with patterned nanogels. A new capillary electrophoresis-mass spectrometry
interface based on acousto-mechanical energy is introduced to enable coupling both techniques without
concern for voltage or flow rate. Aim 1 creates a new functional screening tool for enzyme inhibition
and reduces both the amount of enzyme and the time to evaluate a neuraminidase preparation. The
biocompatibility, automation, and low reagent and sample requirements are harnessed in Aim 2 to
establish a quantitative screening tool to select and evaluate enzyme inhibition of sialylated structures
that interact strongly with the receptor binding domain of the hemagglutinin protein. The full power of
label-free structural identification of capillary electrophoresis interfaced to mass spectrometry outl ined
in Aim 3 leverages the unprecedented gains in signal with electrically assisted vibrational sharp edge
ionization, overcoming barriers of current analytical technologies. The proposed activities are
significant because the low cost, speed, and automation of the separation-based microscale assays
yield previously unattainable information about sialylation fundamental to mitigating viral infections.
These new tools address challenges associated with chemical analyses of sialylated structures to
leverage the role of sialylation in viral infections; thereby, providing researchers the means to combat
virus related diseases and advance human health.

## Key facts

- **NIH application ID:** 10389191
- **Project number:** 3R01GM140560-01S1
- **Recipient organization:** WEST VIRGINIA UNIVERSITY
- **Principal Investigator:** Lisa A Holland
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $205,000
- **Award type:** 3
- **Project period:** 2021-01-01 → 2024-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10389191

## Citation

> US National Institutes of Health, RePORTER application 10389191, Administrative Equipment Supplement to Enabling Technology to Screen and Quantify Sialylated Structures for Activity Against Viral Enzymes and Receptors (3R01GM140560-01S1). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10389191. Licensed CC0.

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