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Introduction toprotein purificationPart 2: protein purification

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Chromatography is the most common technique for purifying proteins

What is chromatography?Separation of components using differential affinities of the components for a mobile phase and for a stationary phase, through which they pass.

“Drawing with colour”(Tswett, inventor of chromatography, 1903)

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Liquid chromatography

• Buffer (mobile phase) pass through a medium (resin) containing chromatography beads (stationary phase)

• Different bead properties can be used to purify different target proteins.– For example, a bead with small

pores can be used to purify a small target from larger molecules in the same sample

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Chromatography principle

Medium

Sample

Target protein

Buffer

Other molecule

s

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Protein properties used in chromatography

Protein property TechniqueBiospecific affinity (tags) Affinity chromatography (AC)Hydrophobicity Hydrophobic interaction

chromatography (HIC)Net charge Ion exchange chromatography

(IEX)Size/volume Size exclusion chromatography

(SEC)(also called gel filtration [GF])

We offer handbooks that are a great reference for practical tips and in-depth information about common chromatography techniques. To download, visit our handbook page

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Schematics of common chromatography techniques

Watch HICprinciple video

Watch IEXprinciple video

Watch SEC (or GF) principle video

Watch ACprinciple video

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Comparison: manual vs automated purificationManual purificationLittle training or start-up time requiredEasy to do parallel runs for increased throughput

Automated purificationAutomation—convenienceGradient elution—high resolutionReproducibilityDocumentation

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Summary protein purification

Efficient purification of proteins is required both in research and in production of biopharmaceuticals.Chromatography is a technique to purify proteins based on properties like size and net charge.Protein purification can be performed manually or automatically.

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Learn more on proteins.gelifesciences.com

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