Application Note: Innovative SPR Assay Transfer to Single Gradient Injections Creating Comparable Kinetics and Affinity Results to Time Consuming Multi-Cycle Kinetics

Overview

Standard multi-cycle kinetics (MCK) and single-cycle kinetics (SCK) require multiple fixed concentrations of an analyte to be injected across a ligand immobilized on a sensor chip surface. This is a time-consuming process, requiring more reagent and setup time for the additional dilutions.

Based on the Taylor dispersion theory, novel OneStep^® injections diffuse a single concentration of analyte into a moving stream of buffer to create an analyte concentration gradient of at least three orders of magnitude. This allows for the accurate measurement of molecular kinetics and affinity from a single analyte concentration. Hence, it takes fewer steps and ligand regenerations to generate the right curvature on the sensogram for kinetics studies increasing sample throughput and kinetic data content, while decreasing sample preparation time and risk of error from preparing multiple sample dilutions. Using a single sensor chip and fewer reagents also lowers assay costs significantly.

Here, a capture-based multi-cycle kinetics assay was independently designed and qualified on an SPR system to measure programmed cell death protein (PD-1) binding to the IgG4 antibody OPDIVO^®. The assay was directly transferred to the novel gradient injection which significantly shortened assay times and achieved comparable kinetics and affinity results to the alternative SPR system.

• Document type: Application Note
• Page count: 7
• Read time: 13 minutes

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