Beyond Sensitivity: The Power of In-Line Mixing for Competitive Compound Analysis
Authors: Stuart Knowling, Ph.D, Sartorius | Last updated: December 2023
Detection and characterization of low molecular weight binding events requires the use of sensitive biophysical techniques that can detect low affinity interactions. Traditionally, surface plasmon resonance (SPR) has been a key technique in screening of these low affinity interactions but the use a single fixed concentration injections can result in the need to make decisions based on inappropriate (small, square-shaped) sensorgrams.
The application note details the application of NeXtStep™ gradient injections using the Octet® SF3 SPR system for competitive compound analysis in drug discovery. Unlike traditional surface competition assays that require the pre-mixing of two compounds, this novel approach offers walk away in-line mixing of compounds for competition assays, meaning multiple combinations of compounds can be easily tested and a competition affinity determined. NeXtStep™ injections are particularly effective for examining the influence of competitor molecules on control molecule binding and for determining the site specificity of small molecules, thereby improving SPR competition assays beyond mere sensitivity.
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Pharmaceutical researchers, biochemists, and drug development professionals involved in competitive compound analysis would benefit from this document. It is also relevant for those looking to improve the efficiency of surface plasmon resonance (SPR) assays in the context of drug discovery.
Figure 1. Cover of application note.
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