Methods of Protein Purification and CharacterizationOBJECTIVES • What characteristics do biomolecules, in general, and proteins, specifically, have that can be used for separation?• Be able to describe the rationale and basic chemistry (not the details) behind the various purification strategies • Be able to design a purification strategy for a mixture of proteins •Given a purification protocol, correctly analyze that strategy and make recommendations for improvement.• What differentiates preparative techniques from analytical techniques? • Be able to determine the sequence of a protein given information from Edman degradation chemistry and proteolytic and chemical fragmentation of the protein.• Why is it useful to know the sequence of a protein?OUTLINEOverview of PurificationWhat characteristics do proteins have that can be used for separation? Measuring the purification processOverview of characterizationStructural FunctionalSeparation on the basis of size/mass:Dialysis Gel Filtration Centrifugation Gel Electrophoresis Separation on the basis of charge Ion exchange chromatography Isoelectric focusing
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