Affinity chromatography is one of the purification and separation methods that have been widely recognized in recent years and have been rapidly developed. Many substances have the property of specifically reversibly binding to a compound. For example: enzymes and coenzymes or enzymes and substrates (products or competitive inhibitors, etc.), antigens and antibodies, lectins and receptors, vitamins and binding proteins, lectins and polysaccharides (or glycoproteins, cell surface receptors), Nucleic acids and complementary strands (or histones, nucleic acid polymerases, binding proteins) and cell and cell surface specific proteins (or lectins). Affinity chromatography is a method in which a compound (called a ligand) which can reversibly specifically bind to a substance to be separated is chemically coupled to a solid phase carrier, and a solid phase carrier carrying a ligand is packed in a column. When the purified biomacromolecules pass through the column, the biomacromolecules specifically bind to the ligand on the carrier and remain on the column, and other substances are washed out. Then, the biomacromolecule is separated and eluted from the ligand by an appropriate method to achieve separation and purification (Fig. 3-10). Shanghai Chuangsai Technology provides 9008-97-3, Phytohemagglutinin BR, product number: D23-RD1047-1g, price 560 yuan.
Affinity chromatography is the most ideal purification and concentration method because it is a ligand and a substance to be separated for specific binding, so the separation and purification efficiency is extremely high, and the purity can be increased by several thousand times. The affinity binding property of the ligand of the affinity chromatography to the substance to be separated can also be used to extract the undenatured portion from the denatured sample, purify a small amount of the desired component from a large amount of contaminated material, and affinity chromatography can also be used. To concentrate its solute from extremely thin liquids. Shanghai Chuangsai Technology Co., Ltd. provides Sephadex G-50 medium, Sephadex G-50 medium, product number: B11000414-25g, price 504 yuan.
The carrier used for affinity chromatography and gel filtration have the same gel properties, that is, chemically stable, non-charged, weak in adsorption capacity, loose in mesh, good in mechanical strength, not easily deformed, and capable of ensuring flow rate. Polyacrylamide gel particles, dextran gel particles, and agarose gel particles are all available, with the agarose gel (Sepharose 4B) type being the most widely used. The key to affinity chromatography is to try to select the appropriate ligand and chemically link the ligand to the carrier to form a stable covalent bond, which needs to be selected and tested as needed in the actual work. Shanghai Chuangsai Technology provides 61970-08-9, agarose gel CL-4B, BR, product number: D16-1027316-100ml, the price is 460 yuan.
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