TY - UNPB
T1 - Observing picomolar protein unfolding using resonance light scattering
AU - Alvarez Bolano, Alain
AU - Arvesen, Kristian B.
AU - Hjuler, Kasper F.
AU - Bjerring, Peter
AU - Petersen, Steffen B.
PY - 2024/11/22
Y1 - 2024/11/22
N2 - 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 seconds. 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 the approximately 1000-fold more concentrated HSA.
AB - 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 seconds. 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 the approximately 1000-fold more concentrated HSA.
U2 - 10.1101/2024.11.20.624557
DO - 10.1101/2024.11.20.624557
M3 - Preprint
BT - Observing picomolar protein unfolding using resonance light scattering
PB - bioRxiv
ER -