Discussion
Which one of the following methods separates biomolecules based on their hydrodynamic volumes?
Answer: C
The method is size-exclusion chromatography.
Size-exclusion (gel filtration) chromatography uses beads with pores of defined size. Molecules with a small hydrodynamic volume enter the pores and take a longer path, so they elute late; large molecules are excluded and come out first. Separation therefore depends on effective size and shape in solution.
Anion- and cation-exchange chromatography separate by charge, binding molecules to oppositely charged groups and eluting them with salt or pH gradients. Thin-layer chromatography separates by differential adsorption and partition between a stationary layer and a moving solvent, essentially by polarity.
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