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Enzyme Purification by Ion Exchange Chromatography

Creative Enzymes has developed versatile purification techniques to satisfy both research demands and the industries applications. We demonstrated several enzyme purification methods to obtain more concentrated and purified enzymes after production. The purification services are of high-quality, fast turnaround, and cost efficiency. Our research center is equipped with cutting-edge instruments and specialized crews. With the professional knowledge and practice of the ion exchange chromatography (IEC) Creative Enzymes provides enzyme purification services.

Ion exchange has been proven to be one of the major methods of fractionation of labile biological substances. With the development of modern high performance media for purification, IEC plays an increasingly important role in the separation and purification of biomolecules. This developed method makes great contribution to the understanding of biological processes, efficient isolation, and getting improved grade products. Normally the isoforms of an enzyme have approximately the same molecular weight, which makes the separation impossible by gel filtration. However, the small differences in charge properties resulting from altered amino acid composition enable the separation of isoenzymes using ion exchange chromatography.

The isoelectric point (pI) is the pH at which the net charge of an enzyme is zero. Arginine, lysine and histidine contribute to positive charges, depending on the pH of the surrounding buffer. Any free N-terminal amine will also contribute to a positive charge below pH 8. Usually, aspartate, glutamate residues, and the C-terminal carboxyl group contribute to negative charges. Therefore, it is possible to separate enzymes using either fixed positive charges on the stationary phase, anion exchanger, or fixed negative charges, cation exchanger. Generally, anion exchange chromatography (AIEXC) is carried out at pH values above the isoelectric point of the enzyme of interest, while cation exchange chromatography (CIEXC) is carried out below the isoelectric point, attracting positively charged enzymes. By simply changing the pH of elution buffer, the protein of interest can be collected.

Enzyme Purification by Ion Exchange Chromatography Figure: The principle of ion exchange chromatography.

The basic isolation principle of IEC is simple but efficient, which make IEC develops to be the widely used approach for the purification of enzymes and proteins. With the exceptionally strong technical development in the application of ion exchange chromatography Creative Enzymes has accomplished thousands of purification projects. If you are seeking for a reliable IEC purification service, Creative Enzymes is the top choice with unique advantages:

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