Structural Determinants for the Interactions of Chemically Modified Nucleic Acids with the Stabilin-2 Clearance Receptor

Hans Gaus, Colton M. Miller, Punit P. Seth, Edward N. Harris

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

The Stabilin receptors are systemic clearance receptors for some classes of chemically modified nucleic acid therapeutics. In this study, the recombinant human secreted ecto-domain of the small isoform of Stabilin-2 (s190) was purified from cell culture and evaluated for direct binding with a multitude of antisense oligonucleotides (ASOs) using a fluorescence polarization-based assay. The tested ASOs varied in their backbone composition, modification of the ribose 2′ position, overall length of the oligo, and sequence of the nucleotide bases. A fully phosphorothioate (PS) ASO with a 5-10-5 pattern of flanking 2′-O-methoxyethyl modifications was then used to test the effects of pH and salt concentration on receptor binding. These tests concluded that the PS backbone was the primary determinant for ASO binding and that decreasing pH and increasing salt generally increased the rate of ligand dissociation and fit within the biological parameters expected of a constitutive recycling receptor. These results will be useful in the rational design of therapeutic oligonucleotides for enhancing their affinity or avoidance of the Stabilin receptors.

Original languageEnglish (US)
Pages (from-to)2061-2064
Number of pages4
JournalBiochemistry
Volume57
Issue number14
DOIs
StatePublished - Apr 10 2018

Fingerprint

Antisense Oligonucleotides
Nucleic Acids
Salts
Phosphorothioate Oligonucleotides
Fluorescence Polarization
Ribose
Recycling
Cell culture
Oligonucleotides
Assays
Protein Isoforms
Nucleotides
Cell Culture Techniques
Fluorescence
Polarization
Ligands
Therapeutics
Chemical analysis

ASJC Scopus subject areas

  • Biochemistry

Cite this

Structural Determinants for the Interactions of Chemically Modified Nucleic Acids with the Stabilin-2 Clearance Receptor. / Gaus, Hans; Miller, Colton M.; Seth, Punit P.; Harris, Edward N.

In: Biochemistry, Vol. 57, No. 14, 10.04.2018, p. 2061-2064.

Research output: Contribution to journalArticle

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