Organic solvent extraction for protein purification
Protein separation purification is widely used in the research and application of biochemistry and is an important operational skill. SCG Protein Purification System Co., Ltd. has organized the organic solvent extraction content for protein purification for everyone. A typical eukaryotic cell can contain thousands of different proteins, some are very rich, and some contain only a few copies. In order to study a certain protein, it is necessary to first purify the protein from other protein and non-protein molecules.
Protein purification ion exchange chromatography with reagents and ion exchangers When the separate protein solutions flow through the ion exchange chromatography column, the protein with the opposite charge of the ion exchanger is adsorbed on the ion exchanger, and then the pH is changed Or method to elute the adsorbed protein.
Organic solvent extraction
The principle of protein purification organic solvent extraction is: organic solvents that are miscible with water (such as methanol and ethanol) can significantly reduce the solubility of some proteins in water; and at a certain temperature, pH and ionic strength, the organic solvent that causes protein deposition The concentration is different, therefore, the protein concentration can be purified separately by controlling the concentration of the organic solvent. For example, under magnetic stirring in an ice bath, slowly participate in ethanol (-25 ℃) in the pre-cooled culture fluid at 4 ℃, the ice nucleoprotein can be separated, and then purified ice nucleoprotein. Because at room temperature, organic solvents can not only cause protein deposition, but also accompanied by denaturation.
Therefore, it is generally necessary to cool the organic solvent, and then participate in the organic solvent under continuous stirring to prevent part of the concentration is too high, the problem of protein denaturation can be largely solved. Some proteins that are more robust to lipids or have more polar side chains in the molecule and are insoluble in water can be extracted with organic solvents such as ethanol, acetone, and butanol.They have certain hydrophilicity and strong lipophilicity. , Is the extract of ambition.
Separate cold ethanol extraction of immunoglobulins was first proposed by Cohn in 1949 and used to prepare gamma globulin. The cold ethanol method is also a method recommended by WHO regulations and biological products regulations in China. It not only has high resolution, good purification effect, but also can separate multiple components together, and has the functions of bacteriostasis, eradication and virus elimination.
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