Chromatography protein purification

Besides protein microarray proteome profiling, the term chemical proteomics (also chemoproteomics or pull-downs ) is mostly used in reference to the application of affinity chromatography protein purification when small molecules are the bait, and liquid chromatography separation of peptides precludes mass spectrometry as the universal readout (LC-MS/MS). 

Reversed-phase chromatography is widely used as an analytical tool for protein chromatography, but it is not as commonly found on a process scale for protein purification because the solvents which make up the mobile phase, ie, acetonitrile, isopropanol, methanol, and ethanol, reversibly or irreversibly denature proteins. Hydrophobic interaction chromatography appears to be the least common process chromatography tool, possibly owing to the relatively high costs of the salts used to make up the mobile phases. 

Before the era of artificially recombinant DNA, affinity chromatography emerged as a potentially highly selective approach to protein purification [13]. It revived a laborious art based on selective precipitations and a limited range of chromatographic media. 

Cuatrecasas, P. (1970). Protein purification by affinity chromatography. Derivatizations of agarose and polyacrylamide beads. J. Biol. Chem. 245, 3059-3065. 

Porath, J. (1992) Immobilized metal ion affinity chromatography. Protein Expr. Purif. 3, 263-281. 

Liquid chromatography is the core preparative technique in protein purification, and all supplementary procedures like extraction, centrifugation and filtration, ultimately serve to condition the protein solution for chromatography. A series of chromatographic steps, usually termed capture, intermediate purification and polishing, mak-... 

Keywords. Short monolithic columns, Monoliths, Chromatography, Separation, Purification, Proteins, DNA, Bioconversion, Solid-phase synthesis... 

Table 10.3 contains a listing of some important proteins. Protein purification must be done under conditions in which conformational and configurational changes are minimal. Such purification is most often carried out using varieties of chromatography including affinity chromatography and electrophoresis. Some of the common features of enzymes are ... 

Procion Rubine MX-B, Procion Yellow H-A, and Turquoise MX-G. These dyes have found use over the last few decades in the purification of a broad range of proteins and enzymes, including albumin, decarboxylases, glycolytic enzymes, hydrolases, lyases, nucleases, oxidoreductases, synthetases, and transferases [77,78], The first use of dye-ligand affinity chromatography was described by Staal et al. in 1971 [79], Since that time, it has become an extremely popular tool for enzyme and protein purification, with hundreds of such compounds having been isolated by this technique [3-6,76-79],... 

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