Notes

Binding Kinetics of Complementary Oligonucleotide Sequences using OpenSPR™

Overview

Oligonucleotides are an important area of biotechnological and pharmaceutical research, as demonstrated by the recent success of mRNA-based COVID-19 vaccines. Oligonucleotides, namely siRNA and RNAi, are also used to treat a number of diseases related to gene expression, due the ability of complementary oligonucleotides to block other molecules from accessing relevant portions of a DNA or mRNA sequence.

Characterizing such molecules is critical for the continued development of these important vaccines and therapeutics. Surface plasmon resonance (SPR) is a well-suited technique for analyzing nucleic acid binding interactions as it can accurately and precisely measure affinities and on/off-rates. In this application note, OpenSPR was used to measure binding of a biotinylated DNA sequence to another DNA sequence containing the reverse complement.

Binding of S-oligo at 200 nM, 67 nM, 22 nM, 7.5 nM and 2.4 nM to the immobilized Bt-oligo. The solid black lines represent the one-to-one diffusion corrected kinetic model fits.