Simplified Protein Purification Using Protein-Polysaccharide Conjugation

dc.contributor.advisorBentley, William Een_US
dc.contributor.advisorEhrman, Sheryl Hen_US
dc.contributor.advisorRaghavan, Srinivasa Ren_US
dc.contributor.advisorWeigand, William Aen_US
dc.contributor.authorSmall, David Andrewen_US
dc.contributor.departmentChemical Engineeringen_US
dc.date.accessioned2004-05-31T19:39:23Z
dc.date.available2004-05-31T19:39:23Z
dc.date.issued2003-12-05en_US
dc.description.abstractThe purification step of protein production and isolation is the most time consuming and complex step in processing. Optimization of the purification process requires rapid, economical, and cost-effective methods to purify proteins at both the small-scale and the large-scale. Instead of using complicated chromatographic techniques and dialysis, a more simplified method is proposed utilizing only a centrifuge. A tyrosine tag with an enterokinase enzymatic cleavage site was added to the C-terminus of green fluorescent protein (GFP). The tyrosine tagged protein was expressed in E. coli cells. The cells were harvested and lysed. The lysate containing the tyrosine tagged GFP was covalently coupled to a polysaccharide (chitosan) using tyrosinase enzyme. The GFP in the protein-polysaccharide conjugate was then liberated using the enzyme, enterokinase. This method was effective in conjugating and purifying the protein using a centrifuge with a limited number of processing steps.en_US
dc.format.extent1360826 bytes
dc.format.mimetypeapplication/pdf
dc.identifier.urihttp://hdl.handle.net/1903/68
dc.language.isoen_US
dc.relation.isAvailableAtDigital Repository at the University of Marylanden_US
dc.relation.isAvailableAtUniversity of Maryland (College Park, Md.)en_US
dc.subject.pqcontrolledEngineering, Chemicalen_US
dc.titleSimplified Protein Purification Using Protein-Polysaccharide Conjugationen_US
dc.typeThesisen_US

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