Question

to compare physical differences between four isoforms of proteins you have to isolate them. describe an affinity coloumn for this purification.

Answer 1

Ans)

Proteins can be separated on the basis of size, charge, solubility and binding affinity. To separate proteins there are many ways which are widely used such as dialysis, gel-filtration chromatography, ion exchange chromatography, affinity chromatography and many more. Chromatography being a widely and successful method as a seoaration technique is used for the separation of proteins as well.

Affinity chromatography is a powerful and generally applicable means of purifying proteins. This technique takes advantage of the high affinity of many proteins for specific chemical groups. proteins that regulate gene expression by binding to specific DNA sequences. A protein mixture is percolated through a column containing specific DNA sequences attached to a matrix; proteins with a high affinity for the sequence will bind and be retained. Affinity chromatography is most effective when the interaction of the protein and the molecule that is used as the bait is highly specific.