BSCI-2520 Chapter 5.2: Chapter 5.2
8 Sep 2016
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Protein purification ph, temperature, and other factors must be controlled ph: buffer solutions to prevent denaturation. Many proteins are denatured by contact with certain surfaces (air/water interface, plastic, glass). Therefore, most protein solutions are handled so as to minimize foaming and are kept relatively concentrated. Long-term storage: factors such as slow oxidation and microbial contamination must be considered. Protein solutions are often stored under nitrogen or argon gas and/or are frozen at liquid nitrogen temperatures. Purification requires some way to quantitatively detect the protein. For enzymes with readily detectable products can be detected by adding the reactants and measuring the rate of product formation. This is proportional to the amount of enzyme present. For non-readily detectable products, another enzyme can be used to convert the product to something that is more easily detected. Proteins that are not enzymes can be detected by their ability to bind specific substances or to produce observable biological effects.
