应用基因工程方法制备卵巢癌抗OC_（125）独特型抗体  

GenerationofGeneticEngineeringAnti┐idiotypicAntibodyAgainstOC125MonoclonalAntibodyofOvarianCarcinoma

作　　者：张香云[1,2] 崔恒[1,2] 钱和年[1,2] 冯捷 邱并生[1,2] 

机构地区：[1]北京医科大学人民医院妇科肿瘤中心 [2]中国科学院微生物研究所

出　　处：《中华妇产科杂志》1996年第11期646-648,共3页Chinese Journal of Obstetrics and Gynecology

基　　金：国家教育委员会回国人员启动基金

摘　　要：目的：扩增和表达卵巢癌抗独特型鼠单克隆抗体可变区基因，用于制备新型卵巢癌疫苗。方法：自产生卵巢癌抗原内影象型抗ＯＣ１２５独特型抗体的小鼠杂交瘤细胞中提取ｍＲＮＡ，采用反转录聚合酶链方法，扩增抗体轻、重链可变区基因，连接成单链后重组至表面表达载体ｐＣＡＮＴＡＢ５Ｅ，转化大肠杆菌ＴＧ１，用辅助噬菌体拯救包装成完整噬菌体，用酶联免疫吸附法进行筛选鉴定。结果：获得抗ＯＣ１２５独特型抗体轻、重链可变区基因，长度约３２５ｂｐ和３４０ｂｐ，单链可变区抗体基因长约７５０ｂｐ；获得有表达的阳性克隆，表达产物能与ＯＣ１２５特异结合。结论：应用基因工程的方法。Objectives:Toamplifyandexpressthevariableregiongenesofmurinederivedanti-idiotypicmonoclonalantibodyandtodevelopanewkindofovariancarcinomavaccine.Methods:mR-NAwaspreparedfrommurinehybridomawhichproducesanti-idiotypicantibodyagainstOC125.Theheavyandlightchainvariableregiongenes(VHandVL)wereamplifiedbypolymerasechainreaction(PCR),joinedbyDNAlinkerfragmentstoformsinglechainFvgene(ScFv),andwereligatedintophagemidvector,pCANTAB5E.TherecombinantvectorwasinducedintocompetentE.coliTG1cels.Thephagedisplayedrecombinantantibodiesweredetectedandidentifiedbyenzyme-linkedim-munosorbentassay(ELISA).Results:TheVH,VLandScFvgeneswere340bp,325bpand750bprespectively.Someofthetransformedclonesexpressedengineeredanti-idiotypicantibodies,whichcouldspecificalybindtotheoriginalantibody(OC125).Conclusions:Geneticengineeringanti-idio-typicantibodyagainstOC125monoclonalantibodywassuccessfulygenerated.Clinicalapplicationofanti-idiotypicantibodytoimmunotherapyandtopreventionofovariancarcinomamaybestudied.

关 键 词：卵巢肿瘤 免疫疗法 抗独特型抗体 

分 类 号：R737.310.3[医药卫生—肿瘤]