# Developing nanobody immune libraries against native neuronal nicotinic receptor complexes

> **NIH NIH R03** · UNIVERSITY OF ILLINOIS AT CHICAGO · 2024 · $79,950

## Abstract

ABSTRACT
Nicotinic acetylcholine receptors are ubiquitous in the brain and underlie regulation of attention and arousal by
cholinergic neurons. nAChRs are highly expressed in neurons of the reinforcement and reward pathways, and
maladaptive effects on nAChRs resulting from drug exposure contribute to drug addiction. These receptors are
crucial in addiction both to nicotine (a direct orthosteric agonist of nAChRs) and to other drugs of abuse
resulting from drug induced plastic changes in cholinergic activity in reward centers. A persistent challenge
facing any nAChR-targeted pharmaceutical approach against addiction is the broad diversity in assembly and
distribution of the brain-expressed forms of the protein. The long-term goal of this project is to develop an
accurate and comprehensive methodology to identify and therapeutically target the full family of native nAChR
conformations. The short-term goal of this R03 proposal is to demonstrate that a yeast-display nanobody
library, screened against two distinct, widely expressed and physiologically critical conformations of nAChRs,
will demonstrate the ability to robustly parse two separate stoichiometries of the nAChR in in vitro fluorescence
and electrophysiology assays. The rationale is to deliver proof-of-concept that this approach can specifically
target individual receptor types, to stimulate further characterization of successfully identified reagents for their
possible functionality in modifying nAChR activity, and ultimately to establish the approach as a feasible
method to study the native expression and assembly properties of the entire family of native nAChRs. This
study includes two related objectives: 1) to develop a nanobody library enriched for yeast-mounted variable
domain modules that potently bind purified 42 nAChRs and 2) to establish an effective counter-screening
assay to deplete stoichiometrically non-specific nanobodies to purify interactors that selectively target two
physiologically relevant, structurally distinct assemblies of 42. This represents an innovative application of
the cutting-edge yeast nanobody screening method against a persistent roadblock in understanding the
maladaptive changes to nicotinic receptor protein expression and distribution. Development and delivery of a
biologic toolbox to study and manipulate all native nAChRs in vivo, as is the overall goal of this research, would
represent a significant biomedical advance to investigate and combat addiction and other human ailments
associated with the cholinergic nervous system.

## Key facts

- **NIH application ID:** 10831953
- **Project number:** 5R03DA055205-02
- **Recipient organization:** UNIVERSITY OF ILLINOIS AT CHICAGO
- **Principal Investigator:** Christian Peters
- **Activity code:** R03 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $79,950
- **Award type:** 5
- **Project period:** 2023-05-01 → 2025-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10831953

## Citation

> US National Institutes of Health, RePORTER application 10831953, Developing nanobody immune libraries against native neuronal nicotinic receptor complexes (5R03DA055205-02). Retrieved via AI Analytics 2026-06-01 from https://api.ai-analytics.org/grant/nih/10831953. Licensed CC0.

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