The high binding capacity for both histidine-tagged and untagged proteins at both research and process scale is an advantage, particularly for those working in the area of drug discovery enabling researchers to optimise with other metal ions for increased selectivity for these proteins.
The use of an uncharged medium for Immobilised Metal Ion Affinity Chromatography allows customers to optimise their purification processes by evaluating the binding of a target protein to various metal ions.
The IMAC Sepharose High Performance and IMAC Sepharose 6 Fast Flow products therefore offers flexibility to all users, and is suitable for industrial applications in which the use of nickel ions may be inappropriate for process scale-up.
His-tagged recombinant proteins are proteins with a special "handle" that uniquely distinguish these proteins from other non-tagged proteins and molecules.
"Customers using the Ni Sepharose 6 Fast Flow and Ni Sepharose High Performance media, pre-charged with Ni2+ ions for purifying histidine-tagged proteins expressed the need for uncharged IMAC Sepharose media to enable them to optimise with other metal ions for increased selectivity for histidine-tagged and untagged proteins," said Allan Simpson, vice president of research and analysis, Protein Separations at GE Healthcare.
"For our process customers, IMAC Sepharose 6 Fast Flow is classified as a GE Healthcare BioProcess medium, giving users access to the full Regulatory Support File that accompanies the company's full line of process media."
The most common protein purification techniques are chromatography and filtration. In chromatography the different substances in a solution are separated in a column - a tube made of plastic, glass or stainless steel.
In the column the solution passes through a bed of small, specially designed particles, or media. The media is designed to separate substances with different properties (electric charge, size, etc.) from each other.
Filtration techniques separate substances in a solution by passing them through semi-permeable membranes under pressure, in hollow fiber or flat sheet formats. Membrane separations are a complementary process to chromatography.
GE Healthcare also makes available the IMAC Sepharose 6 Fast Flow, which is available in prepacked HiTrap (1-ml and 5-ml) as well as HiPrep 16/10 (20-ml) columns.
IMAC Sepharose High Performance is available in prepacked HiTrap (1-ml and 5-ml) columns.