Overview
Currently, thermal approaches like differential scanning fluorimetry (DSF) and differential scanning calorimetry (DSC) are often preferred despite drawbacks like non-equilibrium measurements and thermal alteration of solvent conditions.[3] Chemical denaturation has the advantage of reversible, equilibrated measurement of protein stability. Chemical denaturation uses chaotropic agents (like guanidine hydrochloride or urea) to unfold the protein, and by measuring the fraction of unfolded protein at different denaturant concentrations, the stability of
the protein can be determined.
Utilizing chemical denaturation, the stability of a novel mAb was quickly determined for different buffer solutions at different pH values, using the SUPR-CM fluorescence plate reader. Equilibrium conditions were achieved by letting the prepared microplates incubate for 24 hours, as previously determined for this mAb (data not shown).
