Protein purification involves the isolation of proteins from the source such as microbes, yeast, plants, and mammalian cells, and separating them based on differences in their physical properties. The primary objective of a protein purification scheme is to retain the largest amount of the functional protein while having the fewest contaminants. Hence the purification scheme must be optimized to complete this process in the least number of steps.
Biochemists have devoted effort to find convenient methods for protein purification in case they are aiming to obtain a precise characterization and dive inside the target protein. Proteins as biomicromolecules differ in their chemical and physical properties. These physical properties require the understanding of special techniques for studying, such as mass spectroscopy, gel filtration, ion exchanger chromatography, electrophoresis, and chromatofocusing, while the chemical properties include the composition, covalent and non-covalent bonds, and solubility. These methods, thus depend heavily upon being able to isolate and purify the desired proteins in a way that their physical and chemical properties can be understood, along with their tertiary structures and conformational interactions with ligands and substrates.
Separation techniques for proteins and their basis
Protein purification techniques are an important part to customize or produce proteins having specific properties in a way that they can be applied in various industrial processes. Industry standards of protein purification demand a high level of purity. Several techniques for protein purification are used to reach that required level.
Needless to say, they are extremely crucial to biotechnological and in fact in any life science research.
This course will cover important techniques such as electrophoresis, chromatography, etc that are commonly used in protein purification and discusses the advantages, disadvantages, and potential problems of each. Various protein estimation techniques are also covered along with these.