The Profinity eXact fusion-tag system is an E. coli–based expression and purification system that generates a tag-free protein containing its native N-terminal amino acid sequence in a single step.
SDS-PAGE analysis of a purification of Profinity eXact fusions to a 26 kD green fluorescent protein (GFP) and to a 40 kD maltose binding protein (MBP). Crude E. coli lysate containing the expressed proteins (2 ml) was loaded onto a Bio-Scale Mini Profinity eXact 1 ml cartridge on a BioLogic DuoFlow™ system with bind/wash buffer at 1 ml/min. The cartridge was washed with 10 column volumes (CV) of the same buffer at 1 ml/min. Proteins were eluted with 3 CV sodium fluoride buffer at room temperature and at 0.1 ml/min for 30 min. Total purification time required to generate the tag-free proteins without addition of protease was approximately 60 min.
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