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Biochemistry

Learn the toughest concepts covered in Biochemistry with step-by-step video tutorials and practice problems by world-class tutors

5. Protein Techniques

Affinity Chromatography

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Affinity Chromatography

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Problem

In your own words, describe the principles involved in protein purification by affinity chromatography.

Problem

The target protein to be purified is likely eluted from the affinity chromatography column by _______________. Explain potential advantages & disadvantages of the elution strategies.

Altering the pH of the mobile phase.

Addition of a chaotropic agent such as urea.

Addition of salt and/or free ligand.

Raising the temperature in the column.

Problem

A biochemist is attempting to separate a DNA-binding protein (protein X) from other proteins in solution (proteins A, B & C). Consider the chart & answer the questions below about what type of technique is best for separation.

A. What type of chromatography is best for separating protein X from protein A? ___________________________

B. What type of chromatography is best for separating protein X from protein B? ___________________________

C. What type of chromatography is best for separating protein X from protein C? __________________________

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