Protein Purification Service Company（Part ②）
- Ion exchange chromatography
The basis of ion exchange chromatography for the separation of biomolecules is that the material to be separated is oppositely charged with the ion exchanger under certain conditions and thus can compete with it, and different molecules have different types, quantities and distribution of charges under this condition. , Showing the difference in binding strength with the ion exchanger, which is eluted and separated in the order from weak to strong in the ion exchange chromatography. Features of ion exchange chromatography:
①High resolution, with the emergence of various high-efficiency chromatographic media, choosing an appropriate ion exchanger can ensure that ion exchange chromatography has good selectivity and resolution;
② The high protein exchange capacity is conducive to enlarging the scale of separation and its application in industrial production, which is difficult to achieve by methods such as gel filtration;
③ The application is flexible, and the separation process can be optimized by selecting different ion exchange agents, controlling the composition of the buffer, pH, and ionic strength conditions;
④ The separation principle is relatively clear. The technology is based on different charges. However, for large molecules such as proteins, in addition to electrostatic effects, non-ionic interactions such as hydrophobic interactions, hydrogen bonds, and the nature of buffer ions also affect the separation behavior. ;
⑤ The operation is simple and easy. When the sample is separated on a large scale and the resolution requirement is not high, the protein adsorption and desorption can even not be performed in the chromatography column.
- Affinity chromatography
Affinity chromatography uses the biological activity of protein molecules rather than their physical and chemical properties for separation. That is, the specific affinity between protein and ligand is used as the basis for separation, so it is highly selective and its degree of purification can sometimes be Up to 1000 times, so affinity chromatography is a very effective method of protein purification, mostly used to separate a small amount of specific proteins from a large number of complex solutions, and this purification method also has the effect of concentration. Sometimes only one-step separation by affinity chromatography can achieve rapid and satisfactory protein purification, which is unmatched by other purification methods.
The types of binding between proteins and ligands in affinity chromatography include: the active center or allosteric center of the enzyme binds to specific substrates, coenzymes, activators or inhibitors through secondary bonds, antigens and antibodies, hormones, etc. Combination with its receptor, biotin and avidin/streptavidin, glycoprotein and lectin.
- Covalent chromatography
Chromatography techniques used to separate proteins and other biological macromolecules, such as ion exchange chromatography and hydrophobic interaction chromatography, are based on the non-covalent interaction between the molecule to be separated and the adsorbent, while covalent chromatography is The separation or covalent bond formation between the stationary phase and the solute is used to achieve separation.
The chromatographic technique commonly used for biologically active proteins requires that under mild conditions, both stable bonds can be formed and the structure of the immobilized protein can not be destroyed when the immobilized protein is released.
- Electrophoresis technology
Electrophoresis is the movement of charged colloidal particles towards an electrode with opposite electrical properties under the action of an electric field. The separation principle is based on the charge density of biological macromolecules.
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The above is about the protein purification method, protein purification system, protein purification service, protein purification company, etc., from the official website of Medicilon.
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