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using 6. Draw the elution / fractionation profile of all the homogenate proteins

ID: 193754 • Letter: U

Question

using 6. Draw the elution / fractionation profile of all the homogenate proteins separated anion exchange chromatography. List the proteins in order of their elution. Why are these proteins eluted in this order. What is the basis for these proteins being released from the ion exchange column. protein isoelectric protein protein identity wt(kdaltons)point (pl) protein A RNA polymerase protein B alcohol dehydrogenase protein C RNaseA protein D pyruvate kinase protein ERNA synthetase 290 323 98 200 190 9.2 7.7 6.4 4.8 5.0

Explanation / Answer

Anion-exchange chromatography is a process that separates negatively charged substances using an ion exchange resin containing positively charged groups, such as diethyl-aminoethyl groups (DEAE). In Anion exchange chromatiography, the rasin bed is positively charged that binds to negatively charged molecules passing through the chromatography column containing the resin bed. The tightness of the binding between the substance and the resin is based on the strength of the negative charge of the substances. A substance whose pH>pI will have net negative charge whereas, a substance having pH<pI will have net positive charge. And this is the property which is being exploited to perform anion exchange chromatography.

from the list of the protein, elution order will be as follow

protein D, protein E, protein C, Protein B and at last Protein A