111 nmol of an antibody that binds strongly to the antigenprotein G is covalently bound to the support of an affinity columnwith a void volume of 1.54 mL. The association equilibrium constant(KA) for the binding of protein G to the antibody is6.67 × 108 M–1 at pH 7.1. Calculate theretention 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 wouldit take protein G to elute from this column?
tr = ____ min
If a pH 3.2 elution buffer is applied to the column, protein Gelutes at 3.48 min using a flow rate of 2.00 mL/min. Under theseconditions, what is the association equilibrium constant betweenprotein G and the antibody?
KA = ____ M-1
