The Effects of Sample pH and Salinity on KinExA Measurements
Faculty Mentor Information
Daniel Fologea, Boise State University
Presentation Date
7-2025
Abstract
Correct determination of affinity for bi-molecular, reversible interactions is essential for analytical, diagnostic, and scientific purposes. The affinity values reported for myriads of bi-molecular interactions are known to be strongly dependent on methodology adopted for measurements. Nonetheless, quite different affinity values are often reported for identical biological systems and measuring approaches, which question the validity of the measurements. In this line, we hypothesized that the experimental conditions in which the measurements are conducted would also influence the determined affinity values. To demonstrate this hypothesis, we employed a biological test system consisting of Digoxin and their correspondent antibodies and utilized the Kinetics Exclusion Assay (KinExA) technology and instrument to carry out affinity measurements in variable solution conditions. Our results show that both pH and salinity may affect the binding interactions, yet the potential existence of antagonist effects may impede predicting these changes. Understanding the impact of pH and salinity on the measurements KinExA makes is crucial to contextualizing results and maintaining accuracy in the world of kinetics and analytical determinations.
The Effects of Sample pH and Salinity on KinExA Measurements
Correct determination of affinity for bi-molecular, reversible interactions is essential for analytical, diagnostic, and scientific purposes. The affinity values reported for myriads of bi-molecular interactions are known to be strongly dependent on methodology adopted for measurements. Nonetheless, quite different affinity values are often reported for identical biological systems and measuring approaches, which question the validity of the measurements. In this line, we hypothesized that the experimental conditions in which the measurements are conducted would also influence the determined affinity values. To demonstrate this hypothesis, we employed a biological test system consisting of Digoxin and their correspondent antibodies and utilized the Kinetics Exclusion Assay (KinExA) technology and instrument to carry out affinity measurements in variable solution conditions. Our results show that both pH and salinity may affect the binding interactions, yet the potential existence of antagonist effects may impede predicting these changes. Understanding the impact of pH and salinity on the measurements KinExA makes is crucial to contextualizing results and maintaining accuracy in the world of kinetics and analytical determinations.