Protein Purification Workflow Development Using Bio-Rad's NCO' Chromatography System
Application Note Feb 11, 2014
High purity protein samples are essential for applications from protein structure determination and biochemical characterization to antibody production. A standard purification approach is the use of affinity tags such as hexahistidine (6x histidine) or glutathione-S-transferase (GST) tags. These tags increase the throughput and efficiency of the protein purification workflow, as protocols are often readily available or supplied by the manufacturer.
Affinity tagging is not always a viable option. Some proteins are unstable or inactive once tagged or require posttranslational modifications that do not permit recombinant expression. In these cases, researchers often settle for lower purity protein rather than exhaustively explore purification options, since the purification optimization process can be time and labor intensive when no particular column resins or buffer conditions are dictated by an affinity tag.