Soluble Expression, Purification and Characterization of Single-chain Fv Catalytic Antibody(sFv-2F3)

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摘要 TofindoptimalconditionsforexpressingthesolubleformofsFv-2F3andtostudythepurificationandpropertyofitsderivativeSe-sFv-2F3,thepreferredexpressionconditionswereinvestigatedbymeansoforthogonaldesign.Thesecultureconditionsincludedincubationtemperature,inducerconcentration,inductiontimeandcellconcentration.TheevaluationofexpressionwasaccomplishedbytheanalysisofwholecelllysatesandtheyieldofsolublesFv-2F3wascalculatedaccordingtotheanalysisofProfinder(FTI-500,Pharmacia).Thepurificationprocedurewascarriedoutviaatwo-steppurificationprocedureconsistingofion-exchangechromatography,followedbyimmobilizedmetalaffinitychromatography(IMAC).TheantioxidantefficacyofSe-sFv-2F3wasdemonstratedbythedeterminationofthecontentofthemainproductoflipidperoxidation,MDA,theviabilityofcellsandtheactivityofLDH.WeobtainedthepreferredcultureconditionstogrowtheengineeredbacteriaandtheprocedureforpreparingsolublesFv-2F3andconfirmedtheantioxidantefficacyofSe-sFv-2F3.
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出版日期 2004年03月13日(中国期刊网平台首次上网日期,不代表论文的发表时间)
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