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Protein purification by size exclusion chromatography (SEC)

1. A globular protein of size 30kDa exists as a monomer and dimer in solution. The most appropriate technique for separation of these two forms of protein is

a: Size exclusion chromatography Explanation

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b: Affinity chromatography Explanation

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c: Ion-exchange chromatography Explanation

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d: Hydrophobic chromatography Explanation

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2. In size exclusion chromatography, molecules are separated on the basis of their

a: Molecular weight Explanation

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b: Hydrodynamic volume Explanation

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c: Charge Explanation

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d: Adsorptive properties Explanation

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3. Effective separation of the protein sample through the size exclusion chromatography column depends on a number of factors

a: Pore size Explanation

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b: Viscosity of sample and buffer Explanation

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c: Packing density of the column Explanation

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d: All of the above Explanation

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4. What do you mean by resolution in size exclusion chromatography?

a: The degree of separation between peaks Explanation

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b: Range of separation of the column Explanation

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c: Efficiency of the column media to produce narrow and sharp peak Explanation

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d: Both (a) and (c) Explanation

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5. In size exclusion chromatography proteins are detected by measuring their absorbance at

a: 260 nm Explanation

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b: 300 nm Explanation

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c: 280 nm Explanation

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d: 500 nm Explanation

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6. Application of size exclusion chromatography are

a: Isolate one or more components from a protein mixture Explanation

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b: Determine molecular weight of the protein Explanation

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c: Buffer exchange and desalting Explanation

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d: All of the above Explanation

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7. Chromatographic media used for size exclusion chromatography are chosen on the basis of

a: Inertness Explanation

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b: Chemical and physical stability Explanation

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c: None of the above Explanation

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d: Both (a) and (b) Explanation

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8. Molecules that do not enter the chromatography matrix are eluted

a: In the void volume Explanation

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b: When one column volume of buffer has passed through the column Explanation

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c: When half column volume of buffer has passed through the column Explanation

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d: None of the above Explanation

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9. Fractionation range of a column matrix means

a: Range of molecular weights that have partial access to the pores of the particle and can be separated Explanation

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b: Size of the molecules that are excluded from the pores Explanation

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c: Molecules that elute in the void volume Explanation

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d: Selectivity of the matrix Explanation

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10. What might be one of the possible reasons behind a blocked column ?

a: Protein precipitation in the column Explanation

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b: Column is not equilibrated properly Explanation

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c: Column is overloaded with sample Explanation

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d: Both (a) and (c) Explanation

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