Which is used as the basis of separation of proteins by affinity chromatography?
A)The net charge and pI of the protein at the pH of the column.
B)The protein's molecular weight.
C)The protein's density.
D)The selective binding of the protein to a ligand on the column matrix.

Question

Quizplus · Accepted Answer

Affinity chromatography is based on the selective binding of the protein to a ligand on the column matrix. A ligand that specifically binds to the protein of interest is covalently linked to the matrix. When the sample containing the protein is passed through the column, the protein of interest binds to the ligand, while the other proteins flow through. The protein of interest can then be eluted by changing the conditions of the column or by using a competing ligand. The net charge and pI of the protein, its molecular weight, or density do not play a significant role in the separation based on affinity chromatography.

Which Is Used as the Basis of Separation of Proteins