title: An overview of enzymatic reagents for the removal of affinity tags.
abstract: Although they are often exploited to facilitate the expression and purification of recombinant proteins, every affinity tag, whether large or small, has the potential to interfere with the structure and function of its fusion partner. For this reason, reliable methods for removing affinity tags are needed. Only enzymes have the requisite specificity to be generally useful reagents for this purpose. In this review, the advantages and disadvantages of some commonly used endo- and exoproteases are discussed in light of the latest information.
authors.affiliation: Protein Engineering Section, Macromolecular Crystallography Laboratory, Center for Cancer Research, National Cancer Institute at Frederick, P.O. Box B, Frederick, MD 21702-1201, USA. waughd@mail.nih.gov
authors.forename: David S
authors.initials: DS
authors.lastname: Waugh
mesh_terms: D000345:Affinity Labels / Q000737:chemistry*; D000818:Animals; D020134:Catalytic Domain; D002460:Cell Line; D010450:Endopeptidases / Q000737:chemistry*; D004789:Enzyme Activation; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D020689:Exopeptidases / Q000737:chemistry*; D005822:Genetic Vectors / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry; D012229:Rhinovirus / Q000201:enzymology; D012995:Solubility; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D000345:Affinity Labels; D011994:Recombinant Proteins; D010450:Endopeptidases; D020689:Exopeptidases