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ID:41403202
大小:2.41 MB
页数:34页
时间:2019-08-24
《蛋白质层析新填料应用与选择》由会员上传分享,免费在线阅读,更多相关内容在行业资料-天天文库。
1、蛋白质层析新填料应用与选择Imaginationatwork.做好分离纯化需要什么?层析设备控制软件技术方案填料生物分子特性Ionexchangechromatography(IEX)离子交换层析净电荷Affinitychromatography(AC)亲和层析生物学亲和性Tag,PTMHydrophobicinteractionchromatographhy(HIC)疏水层析疏水性大小/体积Gelfiltration(GF)凝胶过滤填料组成基架发展史SephadexSepharoseSeparationSep
2、arationPharmaciaPharmaciaDextranAgroseCapto高流速琼脂糖ChemicalmodificationofagaroseSepharose™4FastFlow(Highporosity)OHCH2OHOOMPaSepharose6FastFlowOOHO(Mediumporosity)OOHO8.0R7.0CaptoFamily(Mediumporosity)6.0OCHOHO2OHOO5.0OOOO4.0R3.02.01.00.0IncreasingflowrateCapt
3、o高流速琼脂糖表面修饰表面衍生物减少非特异性吸附提高蛋白载量改善质量传递控制内表面积通过提高内表面积提高载量ConventionalSurfaceProteinPorousparticleGraftedLayerofaHydrophilicPolymer高动态载量和更快的质量传递CaptoQ/SPImpRes--快流速高分辨率填料40μm粒径Capto™ImpResHiScreenCaptoQImpResHiScreenQHPHiScreenQFF≈>>Seetutorialregardingconfident
4、ialitydisclosures.Deleteifnotneeded.Superdex200Increasevs.Superdex200时间更短,分辨率更高,用途更广Superdex200Superdex200IncreaseMatrixCross-linkedagaroseanddextranFractionationrangeMolecularweightsfrom10000to600000Flow/pressureRigidbeadswithhighRegularpropertiesflow/press
5、ureresistanceAveragebeadsize13µm8.6µmpHstabilityRegularusepH3–12Shortterm(CIP)pH1-14Increase小粒径,大改变Superdex200Increase更高的分辨率,更短的纯化时间IncreasedresolutionDecreasedruntime0.5ml/min->48min0.5ml/min->48minSuperdex20010/300GL:0.5ml/min->48min1.5ml/min->16minSuperde
6、x200Increase10/300GL:磁感应蛋白MagRSuperose6Increase专为大蛋白和蛋白复合体纯化设计ProteinA抗体亲和填料MabSelectSuRe(2005)MabSelect(2001)MabSelectXtra(2005)rProtASepharoseFFMabSelectSuReLX(2012)nProtASepharoseFFrmProtASepharoseFF1980’s1990200020082015NaOH进行CIP,清洁更彻底GdnHCl处理磷酸处理40分钟3小时
7、CIP300循环0.1MNaOH处理40分钟3小时在位清洗/消毒消除残留物无需额外处理/QC成本0.5MNaOH有效灭活病毒,降低生物负荷及内毒素IncreasedviralinactivationIncreasedbioburdencontrol10.0OrganismTypeTime15.0C.albicansyeast1hLRVA.nigermold1h0.0HIVBVDVCPVBHVPOLSV-40MLVADVP.aeruginosabacteriagram-1h0.1M0.5MS.aureusb
8、acteriagram+1h1forreductiontobelowdetectionlimitof<3organisms/mLOnaverage,0.5MNaOHresultsin0.5LRVhigherinactivationversus0.5MNaOHeffectivelyinactivates0.1MNaOHbacteria,moldandyeastInactivationof
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