Suitable for gravity flow and batch purification

His-tagged proteins are efficiently purified by a one-step procedure from crude lysates both under denaturing and non-denaturing conditions.

• Tridentate IDA-linker for protein elution with lower imidazole concentrations compared to tetradentate chelators (e.g. NTA)
• Precharged material with four metal types (Cu²⁺, Ni²⁺, Zn²⁺, Co²⁺) as well as metal free agarose
• Two metal loading densities
• Different formats: bulk material, pre-packed columns and spin columns for standard microcentrifuge

Optimize your purification strategy by a flexible choice of metal ion and metal loading density

Based on the HSAP concept, IDA-immobilized Cu²⁺, Ni²⁺, Zn²⁺ and Co²⁺ ions exhibit different affinities & specificities towards histidines [2,3]. Copper (Cu²⁺): Shows the lowest specificity resulting in high target recoveris. Unspecific protein binding is minimized by low density metal loading. Cobalt (Co²⁺): Shows the highest binding specificity resulting in reduced unspecific protein binding. Target loss is minimized by high density metal loading. Nickel (Ni²⁺) and Zinc (Zn²⁺): Show intermediate selectivity. While using Ni²⁺ ions is the standard method, IDA-immobilized Zn²⁺ may prove superior to either immobilized Cu²⁺ and Ni²⁺ ions, as a result of its relatively low binding affinity for E. coli host cell proteins^[4]. Low density metal loading enhances the qualitative purification of recombinant protein but with lower target recoveries. High density metal loading results in greater purification of recombinant proteins. However, unwanted proteins within the sample will also be bound.

Selected References

[1] Porath et al. (1975) Metal chelate affinity chromatography, a new approach to protein fractionation. Nature 258:598.
[2] Gaberc-Porekar et al. (2001) Perspectives of immobilized-metal affinity chromatography. J. Biochem. Biophys. Methods 49:335.
[3] Ueda et al. (2003) Current and prospective applications of metal ion–protein binding. Journal of Chromatography 988:1.
[4] Richard et al. (2000) Design of Affinity Tags for One-Step Protein Purification from Immobilized Zinc Columns. Biotechnol. Prog. 16:86.