Selecting a GST Affinity Chromatography Product

Extracted from GST Gene Fusion System, GE Healthcare, 2014

A variety of affinity chromatography products are available from GE Healthcare that have glutathione immobilized as ligand: Sepharose High Performance (HP), Sepharose 4 Fast Flow (FF), and Sepharose 4B. The Glutathione Sepharose media are available in several formats, ranging from 96-well filter plates, spin columns, and gravity-flow columns to prepacked HiTrap™ and HiPrep™ columns. The media are also available in lab packs (media packs in sizes from 10 ml to 500 ml).

These media vary in their performance parameters, as shown in Table 3.1 and Figure 3.2. As seen in the table, a suitable medium can be selected based on which criteria are most important. For example, for highest capacity and yield choose Glutathione Sepharose 4B, for high eluate concentration choose Glutathione Sepharose HP, and for shortest purification time and for scale-up, choose Glutathione Sepharose FF, which allows a higher flow rate.

Fig 3.2. Twenty ml of clarified E. coli lysate spiked with GST-hippocalcin was loaded on each of three, 1 ml GSTrap columns: GSTrap 4B, GSTrap FF, and GSTrap HP, respectively. The flow rate during sample application was 0.3 ml/min, and during equilibration, wash, and elution it was 1 ml/min.

Fig 3.2. Twenty ml of clarified E. coli lysate spiked with GST-hippocalcin was loaded on each of three, 1 ml GSTrap columns: GSTrap 4B, GSTrap FF, and GSTrap HP, respectively. The flow rate during sample application was 0.3 ml/min, and during equilibration, wash, and elution it was 1 ml/min.

Table 3.1. Purification options for GST-tagged proteins using Glutathione Sepharose products (Click image to enlarge)

Purification options for GST-tagged proteins using Glutathione Sepharose products

Selection Guide Sepharose

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Materials

     
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