Protein Purification Techniques
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Transcript of Protein Purification Techniques
Imagination at work
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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