Binding Specificity Challenges in Biosensor-Based Platforms: Non-specific Binding and Potential Mitigation Factors Article
Last updated: January 2025
Overview
Researchers are increasingly adopting biosensor-based analytical techniques for characterizing the affinity and binding kinetics of biotherapeutics, as well as measuring the amount of active analyte in a given sample. These label-free, real-time kinetic assays streamline workflows and expedite assay development. This document explores the challenges and solutions related to non-specific binding (NSB) in biosensor-based affinity characterization, with a focus on biolayer interferometry (BLI).
NSB can compromise the accuracy of these measurements when the analyte of interest binds non-specifically to materials other than the target molecule, or when other molecules in the sample bind non-specifically to the target protein or sensor. The document reviews the challenges posed by NSB in biosensor-based platforms and outlines a Design of Experiments (DOE) approach that systematically evaluates and screen multiple conditions for their ability to reduce NSB, saving time and resources. Two DOE experiments, utilizing Sartorius MODDE® software, are explored and discussed.
Access the full document to enhance your understanding of NSB and optimize your biosensor assays.
- Document type: Scientific Article
- Page count: 12
- Read time: 15 minutes
Key Takeaways
- Impact on Measurements: NSB can interfere with the accuracy of biosensor assays, by masking true specific binding events, leading to inaccurate kinetic parameter calculations.
- Contributing Factors: The biophysical properties of the analyte of interest in relation to the biosensor or ligand can influence NSB. These properties may include hydrophobicity, structure, or isoelectric point.
- Mitigation Strategies: A variety of conditions and NSB mitigators are provided including the benefits of using Octet® Kinetics Buffer.
- Design of Experiments (DOE): A DOE approach, using Sartorius MODDE® software, enables efficient screening of conditions to reduce NSB, identifying optimal conditions by assessing various mitigators and buffer compositions.
