size exclusion chromatography

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7 Terms

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gel filtration

separated molecules according to size

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gel filtration media

  • porous

  • high mechanical strength

  • inert

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separation by size order of elution

large proteins come out first, small last

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when to use gel filtration

  • group separation

    • desalting

    • comopnenets are separated into two major groups according to size

  • higher resolution fractionation of biomolecules

    • polishing

    • performed at later stage of purification as there might still be many types of proteins of similar sizes

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how much sample volume should be loaded

30% of cv for group separation

higher resolution fractionation: 1-2% cv

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advantages

recoveries are high

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disadvantage

low sample loading in fractionation mode