Proteins can be purified by affinity chromatography. Affinity columns work well

Question

Proteins can be purified by affinity chromatography. Affinity columns work well because of the binding of specific protein(s) to the column containing an immobile: H-bond. ionic bond. non-polar group. legend. all of the above. The proteins selected in question 11 can be eluted from the column by: increasing concentration of free legend. increasing the (NaCI). changing the pH boiling the column, e. none of the above. in molecular sieving chromatography (gel filtration), the smaller molecules: spend most of their time in the gel beads. come out last. remain stuck to the beads. All of the above. Suppose you isolated an nonapeptide from biochemistry lab student's blood. After getting it absolutely pure you acid hydrolyzed it to find 0.4 pmoles each of R, E, H, M, S, T, C, Y, I. This indicates that the peptide contains only one: Glutamate. Tryptophan. Alanine. Thiamine. Glutamine.

Explanation / Answer

Based on the given data,

11)

Proteins can be purified by affinity chromatography by using ligands as tolls for purification. For example, cofactors are used as ligands for separation of enzymes and lectins are used for purification of carbohydrates.   

Thus, the correct option is (d).

12)

The proteins are eluted from the column by using High salt buffers. The High salt buffer binds to positively charged ligands and displaces the negatively charged proteins form ligands. This principle is called charge displacement.

Thus, the correct option is (b).

13)

In gel filtration chromatography the larger proteins elutes first from the column and then the elution of smaller proteins takes place. This is because after addition of crude sample, the smaller proteins are stuck inside the gel and spend more time, but larger proteins cannot. So, in first washing larger proteins elute from column and in second washing smaller proteins will elute.

Thus, the correct option is (d) All of the above.

14)

Here, the peptide has following amino acids:

Thus, the correct option is (a) glutamate.

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