电脑桌面
添加小米粒文库到电脑桌面
安装后可以在桌面快捷访问

抗体噬菌体展示技术VIP免费

抗体噬菌体展示技术_第1页
1/31
抗体噬菌体展示技术_第2页
2/31
抗体噬菌体展示技术_第3页
3/31
AntibodyPhageDisplayMeilingXiong20180629ContentsIntroductionofAbphageDisplayTechnologyAbFormatsforPhageDisplayAbLibrariesConstructionPhageAbSelectionMethods&StrategiesPhageAbScreeningApplicationsInvitroAffinityMaturationExpression&PurificationofPhageAbFragmentsIntroductionofPhageDisplayTechnologyTheFfbacteriophagestructureIntroductionofPhageDisplayTechnologyTheschemeofphagemidvectorIGregion:intergenicregion,usuallycontainsthepackingsequenceandreplicationoriginofminusandplusstrandsMoleculartag:tofacilitatelibraryscreeningandforproteinanalysisRestrictionenzymerecognitionsites:usefulforDNArecombinationandgenemanipulation;multiplecloningsites(MCS)Coatprotein:PIII(largerprotein,lessthan5copies,)PVIII(morethan5copies,decreasedlength)AmbercodonTAG:supEstrains(glutamicacidcodon),non-suppressorstrains(stopcodon)ProteasecleavagesitePromoterSignalpeptides:phageproteintranslocation,crucialfordisplaylevelSelectivemarker:forselectionofinfectedhostcellsIntroductionofPhageDisplayTechnologyNonlyticfilamentousphageisthemostoftenusedforphagedisplay,primarilytheM13andFdstrains.Proteinstobeselectedareinfusedtoallfivecoatproteins,withpIIIandpVIIImostcommonlyused.pIIIproteinisessentialforinfectionofbacteriaHelperphage:wild-typepIIIhelperphageandspecialhelperphageAntigenimmobilizedonmagneticbeads,polystryrenesurfaces,oroncolumns,orisusedinsolutionasbiotinylatedantigenandlatercapturedbyimmobilizedstreptavidinAdvantagesofPhageDisplayforRecombinantAntibodySelectionMoreefficientlythanthroughconventionalhybridomasystem.Cheapertoproducerecombinantantibodiesusingbacteria,ratherthanmammaliancellline.Easiertomaintainandgrowbacterialculturesforrecombinantantibodyproduction.Bypassimmunizationinantibodyselection.Bypasstheuseofanimalcellsforproductionofantibodies.Producingthecombinatoriallibrary(ideallywith108to109members)offunctionalantibodiestogeneratealargerrepertoireofantibodiesthanthoseavailablethroughconventionalhybridomatechnology.Easyisolationandexpressionoftheclonedgeneinabacterialhost.Excellentpotentialtofurtherimprovebindingpropertiesoftheselectedantibodybyproteinengineeringtechniques.Capableofgeneratingantibodiesagainstalmostanydesiredantigen,includinghighlyconservedorself-antigens,conformationalvariants,lowimmunogenicantigens,andalsotoxiccomponents,whichisnotpossiblebyinvivoimmunizationofanimals.Anumberofstartingmaterial:proteins,peptides,haptens,celllines,tissueslides,orvirusparticlesAntibodyFormatsThemostcommonlyusedformat:single-chainvariablefragment(scFv)SimplicityofcloningprocessFastandeasylibrarygenerationAhighdisplayrate(smallproteinsize~25kDa)LessstablethanFabfragmentsTendtoformdimers(canbereducedwithlinkermorethan20aminoacids)AntibodyFormatsFabThelightchain(VL-CL)andtheFd-domain(VH-CH1)oftheheavychainofanantibody.Duringbacterialexpression,thesetwochainsaresynthesizedseparately,andsecretedintotheperiplasmwheretheyfoldtoformheterodimers.FabexhibithigherstabilitythanscFvsPossessbetterPKandPDqualitiesthanscFvsEasiertoconvertintofull-lengthantibodiesClinicalapplications:abciximab,lucentis,cimzia.AntibodyFormatsSingledomainantibodyVHH:VHdomainofcamelidantibody,heavychainsonly,IgNAR(newantigenreceptor):sharkantibody,heavychainsonly,UniqueCDRsAffibodiesAnticalinsDARPinsAvimersAffimersMo...

1、当您付费下载文档后,您只拥有了使用权限,并不意味着购买了版权,文档只能用于自身使用,不得用于其他商业用途(如 [转卖]进行直接盈利或[编辑后售卖]进行间接盈利)。
2、本站所有内容均由合作方或网友上传,本站不对文档的完整性、权威性及其观点立场正确性做任何保证或承诺!文档内容仅供研究参考,付费前请自行鉴别。
3、如文档内容存在违规,或者侵犯商业秘密、侵犯著作权等,请点击“违规举报”。

碎片内容

抗体噬菌体展示技术

您可能关注的文档

确认删除?
VIP
微信客服
  • 扫码咨询
会员Q群
  • 会员专属群点击这里加入QQ群
客服邮箱
回到顶部