Ppt0000005 - Understand basic techniques for protein...

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Understand basic techniques for protein separation – by solubility, size, and adsorption Understand the principle and procedure of amino acid analysis Understand the PDCAAS method for analysis of protein nutritional value
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Research applications Small scale ( a few mg or g) Multiple-step procedures Industrial applications Large scale (Tons) One-step procedures
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One step” for food industry can: o Isoelectric precipitation – soy protein isolate, casein o Alcohol precipitation – soy protein concentrate o Ultra filtration – whey protein o Salt precipitation – enzymes Affinity chromatography – antibodies (medical)
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By solubility Salting out Isoelectric precipitation Solvent parcipitation By adsorption Ion exchange Affinity chromatography By size Dialysis Ultra filtration Size exclusion
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Salts: Low concentration increase the solubility of some proteins Precipitate protein from solution as its concentration is increased. Actions are somewhat unique to each protein. Ammonium sulfate (NH 4 ) 2 SO 4 is commonly used because it is highly soluble and very effective. Google “ammonium sulfate calculator” NaCl or KCl may be also be used to “salt out” proteins.
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below precipitation point –precipitate other protein of interest remains in that necessary to precipitate the protein of interest. Centrifugation recovers
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This document was uploaded on 03/03/2011.

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Ppt0000005 - Understand basic techniques for protein...

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