An antibody was designed for the affinity purification of a protein (affinity chromatography).

The protein to be purified has the following characteristics: Tm = 32C, Hm = 50kcal/mole, Cp = 0.72 kcal/mole. The dissociation constant between the antibody and the folded protein is 10-8 M. The total protein concentration loaded on the column is 10-7M, and the effective antibody concentration in the packed column is 10-6M.

1. What fraction of the total protein on the column would be recovered in the final elution step, if carried out at room temperature (21C)?

2. What is the equilibrium constant for unfolding at 4C? Based on this answer, would this result in an increase in the amount of protein recovered?