Abstract
The histidine‐tag is widely used to facilitate purification of recombinant expressed proteins. For immobilized metal‐ion affinity chromatography (IMAC) the most used metal ion is Ni2+. Also Co2+ has been shown useful in obtaining high purity with good yields of histidine‐tagged proteins. Prepacked HiTrap™ TALON® crude column facilitates scalable, fast and easy protein purification of histidine‐tagged proteins. Unclarified samples can be loaded directly to the column reducing the total purification time which helps to ensure high concentrated pure and intact target proteins with minimal degradation. Two proteins (produced in E. coli and P. pastoris) were purified ending up in >90% purity and yields of 90%. An automated purification of a kinase was done by IMAC and gel filtration. SDS‐PAGE analysis showed that the obtained purity of the kinase was above 90% and the amount purified protein was 3 mg. Further, four purification cycles, without cleaning and Co2+‐recharging between the runs, show that the HiTrap TALON crude columns can be used several times without loosing performance. Purity and yield was unchanged
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