Automated Multicolumn Purification of a Histidine-Tagged Protein
Approximately 40% of recombinant proteins that are purified use a histidine tag for easy capture. This article covers how to automate the purification of histidine-tagged proteins and how purification conditions can be optimized to an automated four-step purification scheme that uses affinity-, ion exchange-, and size-exclusion columns. Using a multistep purification scheme removes the manual steps that cause loss of precious proteins and take more time, like dialysis, collection, and reinjecting samples. The final purification scheme reduces a 3–4-day process to 11.5 h from start to finish, all while improving reproducibility, yield, and comparable purity.