Protein Purification Strategies
The development of generic purification strategies has revolutionized protein purification by offering speed and simplicity at all scales.
Before starting purification, it is important to define your requirements and objectives. These may include defining the required purity, activity, and quantity of the target, keeping steps and handling to a minimum, and removing critical impurities (e.g., proteases). Whether protein purification is performed at microgram and milligram quantities for research or at kilogram to ton quantities for industry, there are proven strategies that can help you apply a systematic approach to purification.
CIPP Purification Strategy. The capture phase reduces the amount of unwanted proteins and concentrates the target protein. Intermediate purification removes most contaminants that are closely related to the target protein. Polishing removes specified contaminants and unwanted forms of the target protein that may have been formed during isolation and purification.
The overall purification power of CIPP depends on exploiting a combination of techniques with independent selectivities. Together, the techniques give a cost efficient and robust production of the target substance. When planning a CIPP strategy, it is important to take into consideration the scale of the purification process and the desired yield and use of the target protein.
Each purification step in multi-stage purification strategy must have a clearly assigned purpose. Increasing the number of purification steps will often decrease the overall protein recovery, increase purification time, and can be detrimental to activity. For most laboratory-scale work, a two- or three-step purification protocol will be sufficient, whereas difficult purifications may require several additional steps.