一維微流控蛋白質(zhì)微珠陣列的制備、優(yōu)化及應(yīng)用.pdf

1、湖南大學(xué)碩士學(xué)位論文一維微流控蛋白質(zhì)微珠陣列的制備、優(yōu)化及應(yīng)用姓名：陳韻晴申請(qǐng)學(xué)位級(jí)別：碩士專(zhuān)業(yè)：分析化學(xué)指導(dǎo)教師：王柯敏20060801AbstractMicrochiptechnologyhasbecomeoneofthemostimponantareasinthefieldofchemistry鋤dlifcsciences0ne—dimensional(1D)micronuidicbeada玎aysarekindsofnovelb

2、iomolecul盯microanalysisplatfo珊thatweproposedThesemicrochipplatfo珊shaveexcellentadv鋤tagesinflexiblearraydesign，e器ynuidiccontrolandlowconsumptionofthes鋤plesandreagentsInthisthesis，basedonthisplatform，modificationoftheantib

3、odiesonthebeadsurface鋤dexperimentaloptimizationhavebeenfurtherinvestigatedandhaVebeenappliedforpfoteinanalysissuccessfullyThisthesisincludesthreemajorpartsasfollows：1DenanIralizationoftheinnerwanofthemicfonuidicchannelsa

4、ndpreparationoftheantibody-modifiedmicrobeadsInourirlvestigation，non—specificpfoteinadsofptiononthei蚰erwallofthePDMSmicro—channelsc鋤besuccessfullyrestraincdViapropermodi6cation0ftheinnerwanInantibody-beadspreparation，the

5、antibOdyimmobilizationwasinfluencedgreatlybyprotectiVeproteinssuchasbovinesefumalbumin(BSA)，whichafecommonlyaddedatahighconcentfationtopreventthecommercialantibodyprOductsfIDmtiterdecreaseAsimplepretreatmentprotocolbased

6、onul仃a-filtmtionwascafriedOutandwasproVedtobeemcientfortheBSArcmoValApproachofproteinAlinkagefortheimproVementofantibodyi咖obilizationwasalsoinvestigated2OptimizationofexperimentalconditionsThispartincludespreparationofto

7、talproteinsamples行omtheculturecens，on-beadimmuno-bindingreaction她dwashstep，andchoiceOfsamplingThe0ptimizedconditionswereinVestigatedandtheresultsshowthattheperfo珊anceofthel—DproteinarrayshasbeenimproVed3Quantitative孤aIys

8、isofproteinexpressionThesensitiVityandlimitofdetectionwasgainedforthedetectionofP53proteinusingthel—DproteinarraysAlso，wedeteminedP53copynumberinaveragesingleA549ceⅡIntheinustrationofdif話(huà)fentialprOteinexpressionanalysis，

9、athree-prOteinprOfilingofP53，c—Mycandp-ActinwasdemonstratedPha珊aceuticalmediateddif＆rentialexpressionsofP53柚dc—Mycinsevefalkindsofcelllineswereinvestigatedandthereliab訂ityofthel-Dprotein甜rayswereverifiedbyWestemblottingI