蛋白纯化策略

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1、GeneratedbyFoxitPDFCreator©FoxitSoftwarehttp://www.foxitsoftware.comForevaluationonly.PurificationStrategies纯化策略ContentIntroduction介绍BeforePurification…纯化前的准备Strategies纯化策略PurificationofrecombinantandnativeProtein纯化实例Howtogetdesiredresolution?如何取得预期的结果？Summary总结1/GE/GeneratedbyFoxitPDFCreator©Fo

2、xitSoftwarehttp://www.foxitsoftware.comForevaluationonly.ContentIntroduction介绍BeforePurification…纯化前的准备Strategies纯化策略PurificationofrecombinantandnativeProtein纯化实例Howtogetdesiredresolution?如何取得预期的结果？Summary总结Principlesofoperationforchromatographytechniques层析原理GelFiltrationIonExchangeHydrophobicRe

3、versedAffinity分子筛离子交换interactionphase亲和疏水层析反相OSOO4/GE/2010-4-132/GE/GeneratedbyFoxitPDFCreator©FoxitSoftwarehttp://www.foxitsoftware.comForevaluationonly.QuestionHowtousetheseseparationtechnologiestoperformaSUCCESSFULpurification?如何运用各种分离纯化技术来实现成功的分离纯化？5/GE/2010-4-13ContentIntroduction介绍BeforePu

4、rification…纯化前的准备Strategies纯化策略PurificationofrecombinantandnativeProtein纯化实例Howtogetdesiredresolution?如何取得预期的结果？Summary总结3/GE/GeneratedbyFoxitPDFCreator©FoxitSoftwarehttp://www.foxitsoftware.comForevaluationonly.目标:开发有效的蛋白质分离纯化工艺•Maintainedbiologicalactivity活性•Sufficientpurityandquantity纯度&规模•Go

5、odeconomy经济性7/GE/2010-4-13设定目标：需要的蛋白量功能用于免疫的结构治疗用质谱研究抗原研究蛋白pgngµgmggkg8/GE/2010-4-134/GE/GeneratedbyFoxitPDFCreator©FoxitSoftwarehttp://www.foxitsoftware.comForevaluationonly.终产品纯度要求–一般原则:Extremelyhigh极高High高Moderate中等•crystallizationfor•therapy•antigenformonoclonalx-raystudies结晶治疗用antibodyprodu

6、ction免疫•N-terminalsequencing•invivostudiesofanunknownproteinPK/PD动物实验N端测序•mostphysical-chemicalcharacterizationmethods鉴定9/GE/2010-4-13选择不同规模、不同功能的系统BIOPROCESSSCALEINCREASEDTHROUGHPUTLABSCALEÄKTAprocess™ÄKTAxpress™ÄKTApilot™ÄKTAprime™plusÄKTApurifier™ÄKTAexplorer™LABSCALELOWTHROUGHPUTSIMPLEPURIFI

7、CATIONADVANCEDPURIFICATION10/GE/2010-4-135/GE/GeneratedbyFoxitPDFCreator©FoxitSoftwarehttp://www.foxitsoftware.comForevaluationonly.开始纯化之前胞外~10proteins样品来源Lipopolysaccharide70Å•原核or真核?70ÅOutermembrane•胞内or胞外?细胞破碎?Peptidoglyc