Steps of affinity purification

Loading of solution containing the substance to be isolated: The solution is usually a protein rich mixture such as antiserum, which is poured into the elution column and allowed to run through the gel, at a controlled rate. Proteins with specific affinity for immobilized ligand shall bind and other proteins will go in flow through. This follows washing of column with buffer to remove all unbound protein (Fig. 3). Some affinity purification procedures are summarized in the table 1

Figure 3. An outline of affinity purification scheme.