Question

111 nmol of an antibody that binds strongly to the antigen protein G is covalently bound to the support of an affinity column with a void volume of 1.54 mL. The association equilibrium constant (K_A) for the binding of protein G to the antibody is 6.67 × 10⁸ M^–1 at pH 7.1. Calculate the retention factor for protein G on this column at pH 7.1.

k = ___

At pH 7.1 and a flow rate of 2.00 mL/min, how many minutes would it take protein G to elute from this column?

t_{r = ____} min

If a pH 3.2 elution buffer is applied to the column, protein G elutes at 3.48 min using a flow rate of 2.00 mL/min. Under these conditions, what is the association equilibrium constant between protein G and the antibody?

K_A = ____ M^-1

Answer 1