動(dòng)物細(xì)胞生產(chǎn)尿激酶原吳慎劍

IntroductionofprourokinasemyocardialinfarctionIschemiacerebralinfarctionpulmonaryembolismthrombusvenousembolism第1頁(yè)/共15頁(yè)Thehistoryofresearch1986–1990:方繼明AchievingthecDNAgeneofprourokinase1993:李鳳知AchievingaCHOcelllineCL-11Gwithhighexpression1993–2011：肖成祖Finishedsemi-worksproductionandclinicanimalexperienceCultivationbycellmicrocarrierwithhighdensityperfusionFourstepsinpurificationAchievedthecountrynewdrugcertificateCHO:中國(guó)倉(cāng)鼠卵巢第2頁(yè)/共15頁(yè)1.ProduceofprourokinasebycellcultivationCultivatingprourokinaseengineeringcellbymicrocarriersystemwithhighdensitystabilityofconsecutivecultivate2.StepsinpurificationPurificationofprourokinaseThevalidationofabilitytoannihilatevirus第3頁(yè)/共15頁(yè)1.1CultivatingprourokinaseengineeringcellbymicrocarriersystemwithhighdensityPerfusion

systemmicrocarrierchoseGlucoseMC-1

聚苯乙烯類(lèi)Biosilon多孔微載體Cytopore第4頁(yè)/共15頁(yè)第5頁(yè)/共15頁(yè)FindingadditivewithlittleorwithoutserumDMEM/F1211G-SG-SEMBIGBIF-5第6頁(yè)/共15頁(yè)第7頁(yè)/共15頁(yè)Useofantibiotictokeepcellfrompollution第8頁(yè)/共15頁(yè)Broadencultivate第9頁(yè)/共15頁(yè)第10頁(yè)/共15頁(yè)第11頁(yè)/共15頁(yè)

Purificationofprourokinase1.CM-徑向陽(yáng)離子交換色譜——MPC微孔玻璃珠吸附色譜——SephacrylS-200HR凝膠色譜——對(duì)氨基苯甲醚親和色譜2.CM-徑向陽(yáng)離子交換色譜——羥甲基灰石離子交換色譜——SephacrylS-200HR凝膠色譜3.HS-陽(yáng)離子交換灌注色譜——SephacrylS-200HR凝膠色譜——對(duì)氨基苯甲醚親和色譜4.StreamlineSP-陽(yáng)離子交換色譜——SephacrylS-200HR凝膠色譜——對(duì)氨基苯甲醚親和色譜——QAE-Sepharose陰離子交換色譜（表13-4）5.抗體親和層析

第12頁(yè)/共15頁(yè)第13頁(yè)/共15頁(yè)The