資源描述：
《抗HCCR單克隆抗體的制備及其鑒定.pdf》由會(huì)員上傳分享，免費(fèi)在線閱讀，更多相關(guān)內(nèi)容在行業(yè)資料-天天文庫(kù)。
1、實(shí)用臨床醫(yī)藥雜志2·6·JournalofClinicalMedicineinPractice?0l2l川l6卷LI3jfJjJ抗HCCR單克隆抗體的制備及其鑒定郝波，林艷，張國(guó)新，施瑞華(南京醫(yī)科大學(xué)第一附屬醫(yī)院消化實(shí)驗(yàn)室，江蘇南京，210029)摘要：目的制備抗人宮頸癌癌基因(HCCR)單克隆抗體(mAb)，并進(jìn)行鑒定。方法以HCCR重紕蛋n免疫IB／c小鼠，取免疫小鼠的脾細(xì)胞和同系小鼠的骨髓瘤細(xì)胞SP2／0進(jìn)行細(xì)胞融合，常規(guī)篩選雜交瘤細(xì)胞。腹水j秀導(dǎo)法制箭抗人tlCCR單克隆抗體，并經(jīng)蛋白A親合層析進(jìn)行純化。用Westernblot和免疫熒光等
2、方法槍測(cè)ttCCRmAb的特異H-剛純化HCCR蛋白進(jìn)行包被，建立間接ELISA方法，檢測(cè)系列稀釋的HCCRmAb，確定其靈敏度。結(jié)果篩選flj2株分泌抗人H(：(：I{m的雜交瘤細(xì)胞株。Westernblot結(jié)果表明抗人HCCR抗體能與HCCR蛋白特異性結(jié)合，免疫熒光結(jié)果示癩HepG2細(xì)胞rf1胞漿和胞膜均呈陽(yáng)性染色。問(wèn)接ELISA方法檢測(cè)HCCRmAb的靈敏度可達(dá)到1g／L。結(jié)論成功刪崳外鑒定了特片性抗人HCCRmAb的雜交瘤細(xì)胞株，為進(jìn)一步深入研究HCCR的生物學(xué)特征和臨床應(yīng)用打下基礎(chǔ)。關(guān)鍵詞：HCCR；單克隆抗體；雜交瘤中圖分類號(hào)：R446．
3、62文獻(xiàn)標(biāo)志碼：A文章編號(hào)：1672—2353(2012)03—0006—04PreparationandidentificationofmonoclonalantibodyagainsthumancervicalcanceroncogeneHAOBo，LINYan，ZHANGGuo—xin，SHIRui—hua(TheFirstHospitalAffiliatedtoNanjingMedicalUniversity，Nanjing，Jiangsu，210029)ABSTRACT：ObjectiveToprepareandidentifymonocl
4、onalantibodyagainsthumancervicalcanceroncogence(HCCR)．MethodsPurifiedrecombinantHCCRproteinwasusedasantigentoimmunizeBALB／cmice．TheisolatedspleencellsfromtheimmunizedmicewerefusedwithSP2／0．MonoclonalantibodiesagainstHCCRwerepreparedbynormalhybridomatechnology．PurifiedmAbwasprep
5、aredbyproteinAchromatography．ToidentifythecharacteristicsofHCCRmAbs，Westernblotandimmunofluorescencemethodwereemployed．TheindirectELISAWasestablishedwiththeporifledrecombinantHCCRprotein．a(chǎn)ndusedtodetectthesensitivityofmAbsagainstHCCR．ResultsTwohybridomacelllineswhichcouldsecret
6、eanti—humanmAbagainstHCCRwereestablishedbyhybridomatechnique．SpecificityofmAbagainstHCCRwasassessedbyWesternblot．Immunofluores—cenceshowedthatHCCRpositivestainingoccurredinthecellhyalomitomeandmembraneofhepaticcarcinomaHepG2．TheindirectELISAwasestablishedwiththepurifiedHCCRprot
7、ein，andwiththisindirectELISA，aslowas1L0ffunctionalanti—humanmAbagainstHCCRcouldbedetected．ConclusionHumanHCCRhybridomacelllinesandtheirsecretedmAbshavebeensuccessfu11v0b—rainedandidentified．whichlaysasolidfoundationtofurtherstudythebiologicMcharacteristicsandclinicalapplication
8、sofHCCR．KEYWolIS：humancervicalcanceroncogence；monoclon
