Abstract
Immobilized metal affinity chromatography (IMAC) with a nickel chelate is widely used for purification and identification of histidine tagged recombinant fusion proteins. One challenge for purification is the time needed to purify a significant amount of HIS-tagged proteins. Fast Protein Liquid Chromatography (FPLC) and other medium pressure systems are often utilized to increase flow rate and significantly decrease the amount of time required for chromatography. Demonstrated here is a one-step purification of Metal Affinity Tagged (MAT) fusion proteins using the HIS-Select High Flow (HF) Cartridges. The cartridges are pre-packed with HIS-Select HF Affinity Resin, allowing for high binding capacity and highly specific purification of histidine-tagged proteins. The HF Affinity Gel is a highly cross-linked, 6% beaded agarose for higher flow rates and mechanical stability under pressure. The non-charged, hydrophilic linkage of the nickel-nitrilotriacetic acid (Ni-NTA analog) chelate group to the agarose prevents non-specific interactions between the resin and native proteins. The performance of the HIS-Select Affinity Gel was confirmed via SDS-PAGE and Western Blot analysis. In addition, it was demonstrated that the HIS-Select HF Cartridge offers high binding capacity and highly specific purification of histidine-tagged proteins. This data clearly demonstrates that the HIS-Select High Flow Cartridge is truly a one-step solution to large volume, high flow purification of histidine-tagged proteins.
Published Version
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