Question

A student wants to make an affinity column to purify their enzyme Z. They need to decide on the property(ies) of their matrix (bead), to which they will attach the arm and the ligand. Choose all of the following correct answers:

Group of answer choices

bead (matrix) should have small pores

bead (matrix) should have large pores

bead (matrix) should be nonpolar

bead (matrix) should be polar

Answer 1

Right choices are:

Bead (matrix) should have large pores

Bead (matrix) should be nonpolar

Explanation:- Affinity chromatography is based upon specific binding of biomolecules ( example- proteins, nucleic acids) to a ligand which is immobilized on matrix. Generally, the matrix is a bead on which ligand remains attached. Sepharose, a bead-form of agarose is an excellent choice as the matrix in most cases.

The matrix should be physically and chemically inert so that it does not influence the specific binding of the biomolecule to the ligand. Therefore bead (matrix) should be nonpolar to prevent polar-polar nonspecific binding activities.

In affinity chromatography, the bead size and porosity are designed to maximize the surface area so that the coupling a ligand and binding of target biomolecules can be maximized. Therefore, Bead (matrix) should have large pores is the right choice.