Question

Consider an experiment where your goal is to isolate Bruton’s tyrosine kinase (BTK) enzyme from a whole‑cell lysate. You have an affinity chromatography column with a tyrosine kinase inhibitor molecule covalently attached to the beads. The tyrosine kinase inhibitor binds and inhibits BTK. As a result of the experiment, you are able to elute BTK from the column, but in a mixture of other tyrosine kinases. Why are tyrosine kinases other than BTK present in the eluate?

The kinase inhibitor has low specificity.

BTK is rapidly degraded during cell lysis.

BTK is inactive in the cell.

The kinase inhibitor has low binding affinity.

Answer 1

The kinase inhibitor has low specificity i.e why other tyrosine kinase are also present other than BTK . If the inhibitor would have high specificity then only BTK will be present in the Elute , high specificity means binding only to a particular type of kinase not to bind with all kind of kinases present in the cell lysate .