Chris Moran, Bio-Rad Laboratories, Inc., 2000 Alfred Nobel Dr., Hercules, CA 94547
Fast and efficient protein purification is essential to explore the structure and function of proteins in the human proteome. Successful protein purification strategies use experimental conditions where proteins of interest are stable and have chromatographic properties different from contaminating proteins. Significant time is often required to optimize chromatography parameters, such as buffer pH and gradient characteristics. Furthermore, purification strategies need to be easily scaled as the amount of protein required increases from milligram to gram quantities.
The new Maximizer buffer blending system, a component of the BioLogic DuoFlow chromatography system, significantly decreases the time and resources required for chromatography method development. With a single set of three salt solutions, the system can perform several chromatography experiments, varying the pH or gradient characteristics, or both, for each run. It is not necessary to prepare separate buffers for each chromatography experiment, and the system performs the experiments unattended. Data from the experiments are easily viewed and compared using the new Trace Compare feature of the BioLogic DuoFlow software.
We were interested in purifying a recombinant protein in milligram quantities, anticipating future scale-up to preparative quantities. The 25 kD protein of interest is overexpressed as an E. coli inclusion body. Information from similar proteins suggested a stability range of pH 6.09.0. The protein was solubilized and