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Direct Analyte Detection by Affinity Chromatography An example of the simplest and most common operating scheme for affinity chromatography is shown in Fig.After these nonretained solutes have been removed, an elution buffer is applied to dissociate the retained analyte; this com- monly involves changing the pH or buffer composition of the mobile phase (to decrease the strength of the analyte- ligand interaction) or adding a competing agent to the mobile phase (to displace the analyte from the ligand).Later, the initial application buffer is reap- plied to the system, and the column is allowed to regen- erate before the next sample injection.1.

Direct Analyte Detection by Affinity Chromatography
An example of the simplest and most common operating scheme for affinity chromatography is shown in Fig. 1. In this scheme, the sample of interest is first injected onto the affinity column under conditions in which the analyte will bind strongly to the immobilized ligand. This is usually done at a pH and ionic strength that mimic the natural environment of the ligand and analyte. Because of the specificity of the analyte-ligand interaction, other solutes in the sample tend to have little or no binding to the ligand and quickly wash from the column. After these nonretained solutes have been removed, an elution buffer is applied to dissociate the retained analyte; this com- monly involves changing the pH or buffer composition of the mobile phase (to decrease the strength of the analyte- ligand interaction) or adding a competing agent to the mobile phase (to displace the analyte from the ligand). As the analyte elutes, it is then detected or collected for further use. Later, the initial application buffer is reap- plied to the system, and the column is allowed to regen- erate before the next sample injection. The overall result is a separation that is selective and easy to perform. It is this feature that makes this format so appealing for solute purification or for the quantification of sample compo- nents.