Method and system for purifying and quantitating proteins using heme fusion tags

摘要

A method is provided for purifying a protein comprising the steps of providing a heme tag with an open coordination site and tagging the recombinant protein of interest with the heme tag. A resin framework is used, wherein a base that binds to heme is immobilized to the resin, and the open coordination site of the heme tag is capable of reversibly binding to the base immobilized to the resin. The tagged protein is reversibly bound to the resin, then eluted from the resin and quantified. The method enables the tagged protein to be tracked during protein expression or purification, and can be used to identify secretion of a protein to the periplasm or to tag proteins in the cytoplasm.

主权项

1. A method for purifying a protein or peptide of interest
providing a heme tag with an open coordination site; tagging the protein or peptide of interest with the heme tag; providing a resin framework, wherein: a base that binds to heme is immobilized to the resin, and the open coordination site of the heme tag is capable of reversibly binding to the base immobilized to the resin; and reversibly binding the tagged protein or peptide of interest to the resin.