Related Experiment Video
Updated: May 29, 2026

Protein Complex Affinity Capture from Cryomilled Mammalian Cells
Published on: December 9, 2016
An overview of enzymatic reagents for the removal of affinity tags
1Protein 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
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.
Related Concept Videos
Tagging and Fusion Proteins
Affinity Chromatography

