九因子层析纯化工艺的具体步骤

PurificationofFactorIX(Processexample)(1)JournalofChromatographyA,844(1999)119-128Title:ProductionofhighlypurifiedclottingfactorIXbycombinationofdifferentchromatographyAuthor:L.Hoffer,etal(Octapharma)CryopoorplasmaChromatographyColumn:FractogelEMDAmino650M35mmLLoading:23Lper1000mLgelFlowrate:100mL/minWashing;20mMsodiumcitratebuffer,pH7.0,containing0.1MNaClElution;20mMsodiumcitratebuffer,pH7.0,containing0.7MNaClWashing:2MNaClSanitization:0.5MNaOHVirusinactivationChromatographyColumn:OctylSepharoseCL-4B26mmID*40mmLoading:TheeluatefromfirstcolumnwasapplieddirectlyFlowrate:10mL/minWashing:20mMsodiumcitratebuffer,pH7.0,containing1MNaClElution:10mMsodiumcitratebuffer,pH7.0and1%TritonX-100Washing:1MNaClSanitization:0.5MNaOHS/Dtreatment(0.3%TnBPwasaddedtotheeluate,27C,6h)SimilarTOYOPEARLresinisPhenyl-650MorPPG-600M.SimilarTOYOPEARLresinisAmino-650M.ButwehavenodataregardtousingAmino-650Masionexchangeresin.IncaseofusingTOYOPEARL,conditionshouldbechecked.ChromatographyColumn:HeparinSepharoseCL-4B16mmID*100mmLoadingbuffer:20mMsodiumcitrate,pH7.0Flowrate:2mL/minWashing;20mMsodiumcitratebuffer,pH7.0,containing0.25MNaClElution;20mMsodiumcitratebuffer,pH7.0,containing0.45MNaClSimilarTOYOPEARLresinisAFHeparinHC-650M.Itisassumedthatsameconditionisapplicable.