UHN UT University of Toronto University Health Network

 

 

A Rapid and Universal Tandem-Purification Strategy for Recombinant Proteins

 

We have recently developed a novel tandem purification tag for recombinant proteins. The tag contains a 6-histidine tag followed by calmodulin (HiCaM) which combines two purification strategies, immobilized metal-affinity and hydrophobic interaction chromatography, in a simple two-step procedure. The HiCaM purification strategy is rapid, makes use of widely available, high capacity, and inexpensive matrices, and therefore represents an excellent approach for small or large scale purification of recombinant proteins. This universal tag could be widely used in industry for the production of: protein therapeutics or pure proteins for structure determination and structural proteomics. These techniques rely on the use of simple purification procedures which yield pure and functional proteins (1).

 

 

Figure 1: The N-terminal HiCaM purification tag combines a (His)6 sequence for IMAC purification followed by a CaM module for HIC purification with phenyl sepharose. By virtue of the thrombin cleavage site (TCS), the target protein can either be cleaved on-column or eluted and subsequently cleaved with thrombin.

 

Selected References

1) McCluskey AJ, Poon GMK, Gariépy J. (2007). A rapid and universal tandem-purification strategy for recombinant proteins. Protein Science. 16:2726-2732. MandP-2007

 

 

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