DNA-Encoded Chemical Diversification of mRNA Display Libraries
Abstract
mRNA display can survey vast peptide populations, but the chemical diversity available to selection remains largely constrained by translation. DNA-encoded libraries possess the complementary advantage: synthetic diversity can be introduced through split-and-pool chemistry, although without the peptide scaffolds and evolutionary framework provided by translation. Here, we combine these approaches by performing DNA-encoded chemical diversification directly on mRNAdisplayed peptides. Successive DNA-barcoding steps were coupled to cysteine alkylation and lysine acylation, allowing each chemical operation to be recorded while preserving the linkage between peptide and encoding nucleic acid. Application of this workflow to a five-residue peptide library generated approximately 20 million nominal chemical structures. Selection against carbonic anhydrase IX preferentially enriched library members encoded for a carboxybenzene sulfonamide modification, including a lysine-containing peptide that was independently resynthesized and shown to bind CAIX with a Kd of 26 nM. These results establish a complete progression from encoded chemical diversification and selection to hit identification, resynthesis, and binding validation, providing a practical framework for the discovery of chemically modified peptide ligands.
Related
- Hybrid Macrocyclic Peptides — Synthetic Strategies to Diversify and Cyclize Peptide Libraries in Phage Display Selections
- A proximity-driven and regioselective peptide bicyclization (PReP-Bicyc) approach for phage display libraries
- Avidity by Design: Scalable Programmed Synthesis of Bivalent DNA-Encoded Libraries for Enhanced Selection Sensitivity
- Chemical topology of lipo-amino fatty acid mRNA carriers triggers distinct lipid bulk phases and endosomal escape mechanisms☆
- N-glycopedia: libraries for native N-glycan structural analysis
BibTeX
@misc{anacleto2026dnaencoded,
title = {{DNA-Encoded Chemical Diversification of mRNA Display Libraries}},
author = {Emily Anacleto and Emilyn B. Aucoin and Kirsten Monroe and Ryan Hili},
howpublished = {ChemRxiv},
year = {2026},
month = sep,
doi = {10.26434/chemrxiv.15009391/v1},
url = {https://chemrxiv.org/doi/full/10.26434/chemrxiv.15009391/v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - DNA-Encoded Chemical Diversification of mRNA Display Libraries
AU - Emily Anacleto
AU - Emilyn B. Aucoin
AU - Kirsten Monroe
AU - Ryan Hili
PY - 2026
DA - 2026/09/23
PB - ChemRxiv
DO - 10.26434/chemrxiv.15009391/v1
UR - https://chemrxiv.org/doi/full/10.26434/chemrxiv.15009391/v1
AB - mRNA display can survey vast peptide populations, but the chemical diversity available to selection remains largely constrained by translation. DNA-encoded libraries possess the complementary advantage: synthetic diversity can be introduced through split-and-pool chemistry, although without the peptide scaffolds and evolutionary framework provided by translation. Here, we combine these approaches by performing DNA-encoded chemical diversification directly on mRNAdisplayed peptides. Successive DNA-barcoding steps were coupled to cysteine alkylation and lysine acylation, allowing each chemical operation to be recorded while preserving the linkage between peptide and encoding nucleic acid. Application of this workflow to a five-residue peptide library generated approximately 20 million nominal chemical structures. Selection against carbonic anhydrase IX preferentially enriched library members encoded for a carboxybenzene sulfonamide modification, including a lysine-containing peptide that was independently resynthesized and shown to bind CAIX with a Kd of 26 nM. These results establish a complete progression from encoded chemical diversification and selection to hit identification, resynthesis, and binding validation, providing a practical framework for the discovery of chemically modified peptide ligands.
ER -