Affinity chromatography 26. Which order of steps would lead to best results with affinity chromatography? The Best Order of Steps for Optimal Results with Affinity Chromatography

For best results with Affinity Chomatography, choose the right order of steps

The powerful tool of affinity chromatography for protein purification allows for separations of specific proteins using binding interactions. To get optimal results with affinity chromatography experiments, you must follow the right order. The first step is to prepare the sample by eliminating any proteins that might interfere with the binding. You can do this by size exclusion or buffering the sample with a buffer (Barker and al. 2016, 2016). The sample must be placed on the column that contains the immobilized partner. The column should then be rinsed in a buffer that doesn’t contain the target protein but does have salts or other components that could interfere with binding. A buffer with high levels of a competing or chaotropic ligand can also be used to remove the target protein.
Researchers can make sure that affinity chromatography experiments produce the most effective results by following these steps. Keep in mind that experiment success is dependent upon the conditions used, as well as the composition and type of buffers. Cont….