# Towards Single-Molecule Protein Sequencing with Nanopores

> **NIH NIH R01** · UNIVERSITY OF WASHINGTON · 2022 · $603,938

## Abstract

Title:
Towards Single-molecule Protein Sequencing with Nanopores
Abstract:
This proposal concerns the development of a nascent single-molecule proteomics technology
based on nanopore sequencing. In recent work by Brinkerhoff et al., we demonstrated a first-of-
kind method for sequentially reading single peptide molecules with a biological nanopore,
detecting single-amino acid substitutions in individual molecules and showing that extremely high
fidelity may be reached through indefinitely many independent re-reads of individual proteins. This
method does not require chemical labeling of amino acids, can be carried out on commercially
available nanopore sequencing hardware, and retains the properties that enabled the success of
nanopore DNA sequencing: low overhead cost, physical rather than chemical sensitivity to small
changes in single molecules, and the flexibility to be re-engineered to target specific sequencing
applications. In this proposal to fund a collaboration of the Gundlach and Dekker laboratories,
both world leaders in nanopore technology development, we aim to develop this promising new
technology into a fully fledged single-molecule proteomics tool. This will involve characterizing
reads of a broad array of protein sequences, optimising core elements of the sequencing
methodology for protein analysis, preparing natural peptides for reading, and determining the
method's applicability to pressing proteomics problems such as post-translational modification
and splice variant detection. Achieving these aims will position the nanopore protein reader for
immediate application to proteomics problems in fundamental biology and clinical medicine.

## Key facts

- **NIH application ID:** 10498335
- **Project number:** 1R01HG012544-01
- **Recipient organization:** UNIVERSITY OF WASHINGTON
- **Principal Investigator:** Cees Dekker
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $603,938
- **Award type:** 1
- **Project period:** 2022-08-30 → 2025-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10498335

## Citation

> US National Institutes of Health, RePORTER application 10498335, Towards Single-Molecule Protein Sequencing with Nanopores (1R01HG012544-01). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10498335. Licensed CC0.

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