Patent ID: 6291192
Filing Date: 2001-09-18
Classification: B01L,C07K,C12Q,C40B,G01N,Y02A

Abstract:
In a recombinant DNA method of producing a protein comprising expressing a protein heterologous to a host, obtaining said protein in an inclusion body in said host, and recovering functional protein from said inclusion body by folding or renaturing said protein under folding or renaturing conditions, the improvement which comprises ranking the efficacies of said multiplicity of different sets of folding or renaturing conditions by a method which comprises(a) placing a sample of said folded or renatured protein in each of a multiplicity of wells in a microplate, wherein said folded or renatured protein has been previously folded or refolded according to one or more of a combination of said multiplicity of different biochemical conditions;(b) simultaneously heating said multiplicity of wells from step (a);(c) measuring in each of said wells a physical change associated with the thermal unfolding of said protein resulting from said heating;(d) generating a thermal unfolding curve for said protein as a function of temperature for each of said wells;(e) comparing each of said unfolding curves in step (d) to (i) each of said other thermal unfolding curves and to (ii) the thermal unfolding curve obtained for said protein in its native form under a reference set of biochemical conditions; and(f) ranking the efficacies of said combinations of different conditions for facilitating the folding or renaturing of said target molecule according to the change in each of said thermal unfolding curves.