MS04-P10 The Durham Screens: Fast Protein Buffer Optimisation through Differential Scanning FluorimetryModern crystallographers are capable of unprecedented speed and throughput in their work, from target purification and crystallisation to diffraction data acquisition and analysis. Protein stability is a key factor in the crystallisation process; samples must be conformationally homogenous and structurally sound over a period of days to form high-quality diffracting crystals. We present The Durham Screens, a set of three complimentary 96-condition screens designed to efficiently identify conditions favourable to protein stability.
Differential Scanning Fluorimetry (DSF, also known as Thermofluor and the Thermal Shift Assay) has rapidly become the go-to method for protein stability analysis due to its high throughput, low cost and versatility. By monitoring the thermal denaturation of a protein sample using either its intrinsic fluorescence or an environmentally-sensitive fluorescent dye, DSF can compare a range of conditions and identify those that confer the greatest thermal stability.
The Durham Screens are designed around three themes: pH, salts and osmolytes. By deconvoluting the contributions of each buffer component to the overall stability of a protein sample, the screens provide valuable insights to optimise purification protocols, protect stored samples and guide rationally-designed crystallisation trials.
Grøftehauge, M. K., Hajizadeh, N. R., Swann, M. J., & Pohl, E. (2015). Protein–ligand interactions investigated by thermal shift assays (TSA) and dual polarization interferometry (DPI). (2015) Acta Crystallographica Section D: Biological Crystallography, 71(1), 36-44.Keywords: Sample optimisation, DSF