Part A: 109 nmol of an antibody that binds strongly to the antigen protein G is

Question

Part A: 109 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 1.07mL. The association equilibrium constant (Ka) for the binding of protein G to the antibody is 6.14x108 M-1 at pH 7.3. Calculate the retention factor for protein G on this column at pH 7.3.

k=______

Part B: At pH 7.3 and a flow rate of 1.00 mL/min, how many minutes would it take protein G to elute from this column?

tr=_____ min

Part C: If a pH 3.1 elution buffer is applied to the column, protein G elutes at 4.86 min using a flow rate of 1.00 mL/min. Under these conditions what is the association equilibrium constant between protein G and the antibody?

KA=______M-1

Map Sapling Learning 109 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 1.07 mL. The association equilibrium constant (KA) for the binding of protein G to the antibody is 6.14 x108 MT at pH 7.3. Calculate the retention factor for protein Gon this column at pH 7.3. Number At pH 7.3 and a flow rate of 1.00 min, how many minutes would it take protein G to ute from this column Number min If a pH 3.1 elution buffer is applied to the column, protein G elutes at 4.86 min using a flow rate of 1.00 mL/min. Under these conditions, what is the association equilibrium constant between protein G and the antibody? Number M Previous Next Save And Exit

Explanation / Answer

A. Formula to calculate the retention factor is: k= (KaXM) / Vm

where K is association equilibrium constant

M is moles of antibody

Vm is void volume

therefore, retention factor, k= (6.14 X 108 X 1.09 X 10-7) / 1.07

= 62.54

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