Virtual Labs

Protein purification by size exclusion chromatography (SEC)

1. Another term for size exclusion chromatography is:

a: Affinity chromatography Explanation

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b: Desalting Explanation

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c: Gel filtration chromatography Explanation

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d: Ion exchange chromatography Explanation

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2. What are the important factors to be considered while preparing buffer for size exclusion chromatography?

a: Buffer must be compatible with protein stability and activity Explanation

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b: Buffer must be degassed Explanation

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c: Buffer must be filtered Explanation

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d: All of the above Explanation

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3. In a mixture of the four proteins listed below, which would elute second in size exclusion chromatography

a: Cytochrome- 13kDa Explanation

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b: Immunoglobulin- 145 kDa Explanation

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c: Ribonuclease- 13.7 kDa Explanation

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d: RNA polymerase – 45 kDa Explanation

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4. Superdex media is made by

a: Cross-linking dextran Explanation

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b: Cross linking dextran with agarose Explanation

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c: Cross-linking porous agarose Explanation

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d: None of the above Explanation

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5. Analytical size exclusion chromatography is used for

a: Desalting Explanation

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b: Check the quality of the sample Explanation

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c: Isolate one or more components from a sample Explanation

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d: All of the above Explanation

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6. The absorbance of which amino acid is predominantly detected by the UV detector in size exclusion chromatography?

a: Histidine Explanation

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b: Proline Explanation

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c: Tryptophan Explanation

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d: Glutamine Explanation

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7. What can be the possible reasons behind protein molecules not eluting as expected?

a: Column is not equilibrated properly Explanation

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b: Protein has precipitated on the column Explanation

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c: Column is overloaded with sample Explanation

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d: All of the above Explanation

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8. What can cause low recovery of protein activity but normal recovery of protein molecules after purification using size exclusion chromatography?

a: Protein molecule is unstable or inactive in the buffer used Explanation

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b: Protein has been degraded by proteases Explanation

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c: Column is not equilibrated properly Explanation

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d: High flow rate of buffer Explanation

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9. The eluted protein fractions after performing size exclusion chromatography can be analyzed using

a: Agarose gel electrophoresis Explanation

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b: EMSA Explanation

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c: SDS PAGE Explanation

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d: None of the above Explanation

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10. Which of the following solution can be used for cleaning size exclusion chromatography column?

a: 1M NaOH Explanation

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b: 30 % Isopropanol Explanation

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c: Both (a) and( b) Explanation

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d: None of the above Explanation

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