hGH單克隆抗體的制備及雙抗體夾心ELISA檢測(cè)法的建立.pdf

1、獨(dú)創(chuàng)性聲明本人聲明所呈交的學(xué)位論文是本人在導(dǎo)師指導(dǎo)下進(jìn)行的研究工作及取得的研究成果。據(jù)我所知，除‘了文中特別加以標(biāo)注和致謝的地方外，論文中不包含其他人已經(jīng)發(fā)表或撰寫過(guò)的研究成果，也不包含為獲側(cè)委鐮及欠喜或其他教育機(jī)構(gòu)的學(xué)位或證書而使用過(guò)的材料。與我一同工作的同志對(duì)本研究所做的任何貢獻(xiàn)均已在論文中作了明確的說(shuō)明并表示謝意。學(xué)位論文作者簽名：fi驂Z簽字日期：扣，1年多月／日學(xué)位論文版權(quán)使用授權(quán)書本學(xué)位論文作者完全了解安諾殳k售有關(guān)保留、使

2、用學(xué)位論文的規(guī)定，有權(quán)保留并向國(guó)家有關(guān)部門或機(jī)構(gòu)送交論文的復(fù)印件和磁盤，允許論文被查閱和借閱。本人授權(quán)空敞睹以將學(xué)位論文的全部或部分內(nèi)容編入有關(guān)數(shù)據(jù)庫(kù)進(jìn)行檢索，可以采用影印、縮印或掃描等復(fù)制手段保存、匯編學(xué)位論文。(保密的學(xué)位論文在解密后適用本授權(quán)書)學(xué)位論文作者簽名：t乩墨寢、簽字日期：≯oIf年石月，日學(xué)位論文作者畢業(yè)去向：工作單位：通訊地址：導(dǎo)師簽名：簽字日期：電話：郵編：Withthedevelopmentofhybridoma

3、techniqueandtheprogressinenzyme1inkedimmunosorbentassay，itispossibletoestablishthebetterWayiIlmeasuringhGHinbloodserumELISAanalysishasSOmanymeritsinsensitivity、specificity、stabilityItiSinexpensiveandfree仔omradioactivecon

4、tamination111issandwichELISAiSanemcientmethodthatcanbeeasilyadaptedtotheautomateddevicesforconfirmationandmonitoringofhGHdeficiencyInthisstudyweimmunizedrabbitswithrecombinanthumanGrowthHormoneItsspleencellsfusedwithSP2／

5、0andthencultivatedineellcultureplateScreeningthehybridomaeelltogetthemonoclonalantibodies(MeAb)TherearetwoMcAbsweobtainednamed3El1、2G9Byanalysisoftiters，Igsubclass，SDS—PAGE，westemblottingthetitersoftheserumsreachl：100and

6、1：107byELlSAmethod，andtheMcAbshavehiehpurityandspecificityIIlthisstudywedevelopedtheDoubleAntibodySandwichELlSAforhGHThismethodusinganantihGHmonoclonalantibody3EllascoatingantibodytecognizeandbindtohGH，thesecondmonoclona

7、lantibody2G9astheantibodylabeledwiththeenzylTlehorseradishperoxidase(HRP)，toachievethedetectionofhGHWedeterminedtheconcentrationoftheantibody，andtheirworkingconditionsAccordingtothevalueofOD490standardcurvedrawnsuccessfu

8、llyAdoubleantibodysandwichELISAassayWasestablishedDetectionrangeis025—20ng／m1sensitivityofO2ng／ml，thenumberofconcentrationhasgoodlinearrelationshipwimOD490values，Theintraassaycoefficientofvariation(CV)wasbetween656％and87

9、％whiletheinterassayCVWasbetween713％and925％Theaveragerecoverywasbetween9806％ThesefeaturesshowthatourassayisanefficientmethodforthedeterminationofhGHItisexpectedtodevelopagrowthhormonedetectionkitthroughfurtherclinicaltria