# Bioconjugations Employing Unnatural Amino Acids

> **NIH NIH R15** · COLLEGE OF WILLIAM AND MARY · 2020 · $324,678

## Abstract

Project Abstract
The ability to prepare well-defined protein/small-molecule/solid-support bioconjugates
possesses significant advantages in the treatment and diagnosis of a variety of
diseases. However, several issues associated with the preparation of the bioconjugates
preclude their widespread application. This proposal aims to address these issues via
the use of unnatural amino acid technologies to rapidly synthesize highly active and well-
defined conjugates. Specifically, a variety of unnatural amino acids will be synthetically
prepared and assessed for their utility in bioconjugations. First, utilizing pre-existing
UAAs, novel bioconjugations will be developed and optimized under physiological
conditions, including the Glaser Hay coupling between terminal alkynes and the [2+2+2]
cyclotrimerization between three alkynyl moieties. Secondly, a set of amino acids with
reactive moieties (alkyne, azide, aminooxy, aldehyde etc) will be prepared to improve
the efficiency of the bioconjugation reactions and prepare multivalent conjugates.
Following the synthesis and in vitro assessment of these novel amino acids, they will be
evaluated for site-specific incorporation into model proteins using evolved aminoacyl
tRNA synthetase/tRNA pairs. Utilizing optimized conditions, both model and medically
relevant proteins will be immobilized onto solid-supports using the technologies to
develop diagnostic protein chips. Finally, the developed technologies can be combined
to produce multivalent bioconjugates that provide numerous therapeutic properties
inaccessible to standard bivalent conjugates. This includes the linking of an antibody
targeting agent to a small molecule drug and a fluorophore to be able to track drug
delivery; however, there are numerous other applications of multivalent conjugates that
can also be envisioned. Ultimately, the methodologies will be transitioned to biologically
relevant systems including ubiquitin binding proteins, esterases, antibodies, and
Cas9/dCas9. The tools developed within the proposed research will not only significantly
advance the fields of therapeutics and diagnostics, but also be extremely useful in the
training of undergraduate researchers towards their future careers within the scientific
arena.

## Key facts

- **NIH application ID:** 10045962
- **Project number:** 2R15GM113203-02
- **Recipient organization:** COLLEGE OF WILLIAM AND MARY
- **Principal Investigator:** Douglas Young
- **Activity code:** R15 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $324,678
- **Award type:** 2
- **Project period:** 2016-04-01 → 2024-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10045962

## Citation

> US National Institutes of Health, RePORTER application 10045962, Bioconjugations Employing Unnatural Amino Acids (2R15GM113203-02). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10045962. Licensed CC0.

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