1 research outputs found
Temperature Gradient Approach for Rapidly Assessing Sensor Binding Kinetics and Thermodynamics
We report a highly resolved approach
for quantitatively measuring
the temperature dependence of molecular binding in a sensor format.
The method is based on surface plasmon resonance (SPR) imaging measurements
made across a spatial temperature gradient. Simultaneous recording
of sensor response over the range of temperatures spanned by the gradient
avoids many of the complications that arise in the analysis of SPR
measurements where temperature is varied. In addition to simplifying
quantitative analysis of binding interactions, the method allows the
temperature dependence of binding to be monitored as a function of
time, and provides a straightforward route for calibrating how temperature
varies across the gradient. Using DNA hybridization as an example,
we show how the gradient approach can be used to measure the temperature
dependence of binding kinetics and thermodynamics (e.g., melt/denaturation
profile) in a single experiment