Metal‐Chelate Affinity Chromatography

AbstractRecombinant proteins engineered to have six consecutive histidine residues on either the amino or carboxy terminus can be purified using a resin containing nickel ions (Ni2+) that have been immobilized by covalently attached nitrilotriacetic acid (NTA). This technique is know as metal‐chelate affinity chromatography and can be performed using either native or denatured protein. This unit presents protocols for expression of histidine‐tail fusion proteins and their purification in either native or denatured form (along with procedures for renaturation by either dialysis or solid‐phase renaturation). Also provided are procedures for analysis of the purified produce and regeneration of the NTA resin.