Question

The first step in preparation of affinity chromatography column is _____________

(a) ligand attachment to matrix

(b) coupling of aromatic amines to matrix

(c) activation process

(d) precipitation

I had been asked this question during an interview.

The query is from Enzymes Purification in chapter Enzymes Purification of Enzyme Technology

Answer 1

The correct choice is (c) activation process

To explain: Activation process is the first step in preparation in affinity chromatography column which involves introduction of reactive groups into the chemically inert polymeric matrix material. The second step is attaching the ligand to the matrix covalently. After the activation, the activated matrix may be coupled to primary aliphatic or aromatic amines. Elution is the process of removing the adsorbed material from the column. This is the last step in affinity chromatography.