Work

A biomolecular sandwich assay to characterize the binding properties of antisense oligonucleotide conjugates

Year: 2026

Type: preprint

Abstract: Antisense oligonucleotides (ASOs) are a treatment modality for genetic diseases. ASOs bind to ribonucleic acid (RNA) with high specificity, with downstream effects that eventually reduce, restore, or modify protein expression through distinct mechanisms. Conjugating ASOs to targeting ligands has emerged as a promising strategy to improve tissue-specific delivery. Here, we develop streamlined assays that assess the binding kinetics of both the ASO and the targeting ligand using biolayer interferometry (BLI). The utility of this biomolecular sandwich binding assay is demonstrated with phosphorodiamidate morpholino oligomers (PMOs) and peptidenucleic acids (PNAs) conjugated to a peptide or protein ligand. We show that this assay can be used to detect intracellular uptake and predict in vitro efficacy. We believe that the methods developed here can accelerate the development of next-generation ASO therapeutics.

Source: ChemRxiv

DOI: 10.26434/chemrxiv.10001912/v1

Authors: Stephen A. Byrne, Sarah Antilla, Julien Couture-Senécal, Nathan W. Dow, Aurélie Rondon, Belinda B. Zhang, Sunit Pal, Michaela Prado, Ava Self, Andrei Loas, Daniel G. Anderson, Bradley L. Pentelute

Institutions: Koch Institute for Integrative Cancer Research At MIT, Massachusetts Institute of Technology, Broad Institute, Harvard University, Harvard–MIT Division of Health Sciences and Technology

Language: English

Topic: RNA Interference and Gene Delivery

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: green

Funder: National Institutes of Health

OpenAlex ID: W7127425267

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