Membrane proteins are responsible for a large variety of tasks in organisms and of particular interesting as drug targets. At the same time, they are notoriously difficult to work with and require a thorough characterization before proceeding with structural studies. Here, we present a biophysical pipeline to characterize membrane proteins focusing on the optimization of stability, aggregation behavior, and homogeneity. The pipeline shown here is built on three biophysical techniques: differential scanning fluorimetry using native protein fluorescence (nano differential scanning fluorimetry), dynamic light scattering, and mass photometry. For each of these techniques, we provide detailed protocols for performing experiments and data analysis.
Keywords: Biophysical characterization; Detergent screening; Differential scanning fluorimetry; Dynamic light scattering; Interferometric scattering microscopy; Mass photometry; Membrane proteins.
© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.