By Terri Thompson & Peter A. Fung
First published May 9, 2016 on www.Biocompare.com
Separating out a protein from a complex mixture is not an easy task. Many hours are spent by bioprocessing scientists trying to purify their favorite protein to homogeneity in order to study the functional properties or to manufacture a large-scale preparation for production or potentially clinical use. Even 1% contaminant in an otherwise highly purified protein preparation, can reek havoc in a functional assay or structural protein study not to mention in the drug discovery process. As many researchers have learned, the purification process is as much of an art as it is science. As with the more common artistic endeavors, protein purification not only relies on protocols and procedures, it brings together intuition, years of practice and trial and error experimentation. While there are different procedures to purify protein, in today’s lab, the method of choice for separating and purifying a target protein of interest from complex mixtures is liquid chromatography. Here we provide some fundamental tips and tricks to preparative chromatography that will yield a pure sample preparation that will advance your protein characterization studies.