Observing Picomolar Protein Unfolding Using Resonance Light Scattering.

ABSTRACT: We here present a novel and sensitive methodology for determining the melting point (MP) of Bovine Serum Albumin (BSA) from micromolar to picomolar concentration levels under label-free conditions. At 1 pM we could model the melting with a sharp Gaussian. However, from the transient state observed during the melting process by using a simple exponential decay model, we determined a time constant of 67 s. We applied this methodology by studying a 3.3 pM sample of a botulinum toxin A (BoNT-A) (stabilized with 2.8 nanomolar denatured Human Serum Albumin (HSA)). We were able to determine the Tm of BoNT-A in the presence of approximately 1000-fold more concentrated HSA. This method enables the detection of protein melting transitions at picomolar concentrations without the use of a fluorescence

SUBMITTER: Bolano Alvarez A 

PROVIDER: S-EPMC12650451 | biostudies-literature | 2025 Nov

REPOSITORIES: biostudies-literature