Question

Please answer in atleast 1 paragraph...

1. Please explain the principle of affinity chromatography and then provide many (two or three) specific examples.

2. Describe the difference b/t step-gradient and the linear gradient. What are the advantages and the disadvantages in each?

Answer 1

Explanation

1. Affinity as the term suggests means, interaction of two species pertaining to biochemical systems and separation based on this interaction. Say interaction between an antigen and an antibody assisted separation, or enymes binding to a specific substrate, or an enzyme binding specific to a particular inhibitor. In this type of chromatography separation one binding component is immobilized on a support and the other binding agent is flown through it. The strength of binding and specificity of the type of binding enables separation process. Applicable in protein separation.

2. In a linear gradient separation we use only one type of concentration throuhout for the separation. Whereas, in case of step gradient we use changing concentrations over time. We get sharper peaks, better separation and resolution in case of step gradient method. The linear gradient is easy and require less sophisticated equipment.