Process Optimization
Basic characterization of the thermal stability of a protein in its native state, or in site-specific or chemically modified forms, is useful in many applications where this knowledge can frame e.g. a particular isolation or HTS screening strategy. Many proteins show pronounced differences in thermal stability depending on solution ion concentration, pH, or additional factors that can affect protein stability.
Effect of construct design on MAPKAP kinase-2 activity, thermodynamic stability and ligand-binding affinity. Kervinen J, et.al. Arch Biochem Biophys. 2006 May 15;449(1-2):47-56. [Thermofluor characterization of protein constructs for structure-based drug design] PMID: 16620770
Enhancing recombinant protein quality and yield by protein stability profiling. Mezzasalma TM, et.al. J Biomol Screen. 2007 Apr;12(3):418-28. [Optimization of protein isolation conditions.] PMID: 17438070
The use of differential scanning fluorimetry to detect ligand interactions that promote protein stability. Niesen FH, et.al. Nat Protoc (2007) 2:2212-2221 [Identification of protein-binding ligands via fluorescence monitoring of thermal unfolding characteristics.] PMID: 17853878
Methods for protein characterization by mass spectrometry, thermal shift (ThermoFluor) assay, and multiangle or static light scattering. Nettleship JE, et.al. Methods Mol Biol (2008) 426:299-318 [High-throughput fluorescence-based monitoring of protein thermal stability and its conditioning by buffers, additives and ligands.] PMID: 18542872
Crystal and solution structure, stability and post-translational modifications of collapsin response mediator protein 2. Majava V, et.al. FEBS J. 2008 Sep;275(18):4583-96. PMID: 18699782
High throughput methods of assessing protein stability and aggregation. Senisterra GA, Finerty PJ, Mol Biosyst (2009) 5:217-223 [Review comparing differential scanning fluorimetry (DSF) using SYPRO Orange dye, differential static light scattering (DSLS), and isothermal denaturation (ITD) for analysis of protein stability and aggregation] PMID: 19225610
High-throughput thermal scanning: a general, rapid dye-binding thermal shift screen for protein engineering. Lavinder JJ, et.al. J Am Chem Soc (2009) 131:3794-3795 [High-throughput analysis of the thermal stability of engineeered variants of Rop protein.] PMID: 19292479
Novel strategies to overcome expression problems encountered with toxic proteins: application to the production of Lac repressor proteins for NMR studies. Romanuka J, et.al. Protein Expr Purif. 2009 Oct;67(2):104-12. PMID: 19460439
Expression, purification, stability optimization and characterization of human Aurora B kinase domain from E. coli. Sheth, PR et.al. Arch. Biochem.Biophys. 2010 Nov 15;503(2):191-201 [Optimization of protein stability using TdCD measurements] PMID: 20699085
Role of the C-terminal domain of the regulatory subunit of AHAS isozyme III: Use of random mutagenesis with in vivo reconstitution (REM-ivrs). Slutzker A, et.al. Biochim Biophys Acta. 2011 Mar;1814(3):449-55. PMID: 21224018
Determination of the volume changes induced by ligand binding to heat shock protein 90 using high-pressure denaturation. Toleikis Z, et.al. Anal Biochem. 2011 Feb 21. PMID: 21345327