Protocol for evaluating protein-polyfluoroalkyl substances in vitro using differential scanning fluorimetry

Abstract

Summary: Per- and polyfluoroalkyl substances (PFAS) are ubiquitous synthetic chemicals that threaten public health, and serum albumin binding of PFAS represents one major variable influencing PFAS toxicokinetics. In this protocol, we describe a differential scanning fluorimetry (DSF) assay suitable for the rapid determination of the relative binding affinities of serum albumin proteins to different PFAS. Herein, we address common experimental challenges related to PFAS solubility constraints, the high background fluorescence of DSF with serum albumins, and the limitations of using DSF-derived dissociation constants (KD) to quantify PFAS-albumin interactions.For complete details on the use and execution of this protocol, please refer to Jackson et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Health Sciences
• High Throughput Screening
• Systems biology
• Environmental sciences