Affinity Chromatography Techniques

Affinity chromatography separates proteins based on reversible interactions between the proteins and a specific ligand bound to the chromatographic stationary phase. Affinity chromatography is highly specific, providing very good resolution, and considerable capacity for analytes of interest. It is the only technique that enables protein purification on the basis of its biological function or individual chemical structure. Active proteins can be separated from denatured or functionally different forms, allowing isolation of pure substances present at low concentrations in large volumes of crude samples, or the removal of specific contaminants. This module introduces the principles of affinity chromatography.

Topics include:

• Principles of affinity chromatography
• Affinity chromatography stationary phases
• Monoclonal antibody titer determination
• Carrying out a mAb titer analysis
• Clean-in-place protocol (CIP)