title: Formulation screening by differential scanning fluorimetry: how often does it work?
abstract: There is strong evidence to suggest that a protein sample needs to be well folded and uniform in order to form protein crystals, and it is accepted knowledge that the formulation can have profound effects on the behaviour of the protein sample. The technique of differential scanning fluorimetry (DSF) is a very accessible method to determine protein stability as a function of the formulation chemistry and the temperature. A diverse set of 252 soluble protein samples was subjected to a standard formulation-screening protocol using DSF. Automated analysis of the DSF results suggest that in over 35% of cases buffer screening significantly increases the stability of the protein sample. Of the 28 standard formulations tested, three stood out as being statistically better than the others: these included a formulation containing the buffer citrate, long known to be `protein friendly'; bis-tris and ADA were also identified as being very useful buffers in protein formulations.
authors.affiliation: Manufacturing Flagship, CSIRO, 343 Royal Parade, Parkville, VIC 3052, Australia.
authors.forename: Marko
authors.initials: M
authors.lastname: Ristic
authors.affiliation: Manufacturing Flagship, CSIRO, 343 Royal Parade, Parkville, VIC 3052, Australia.
authors.forename: Nicholas
authors.initials: N
authors.lastname: Rosa
authors.affiliation: Manufacturing Flagship, CSIRO, 343 Royal Parade, Parkville, VIC 3052, Australia.
authors.forename: Shane A
authors.initials: SA
authors.lastname: Seabrook
authors.affiliation: Manufacturing Flagship, CSIRO, 343 Royal Parade, Parkville, VIC 3052, Australia.
authors.forename: Janet
authors.initials: J
authors.lastname: Newman
mesh_terms: D000818:Animals; D002021:Buffers; D002256:Carbonic Anhydrases / Q000378:metabolism; D002417:Cattle; D005470:Fluorometry / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point
chemical_list: D002021:Buffers; D002256:Carbonic Anhydrases
keywords: Thermofluor; buffer; differential scanning fluorimetry; formulation; protein stability