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时间:2020-05-03
《异常凝血酶DCP基因克隆表达、纯化及单克隆抗体制备与鉴定-论文.pdf》由会员上传分享,免费在线阅读,更多相关内容在行业资料-天天文库。
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2、辱簪^;l化泰恚.世界华人消化杂志2013年12月18日;21(35):3932—3939wcid@wjgnet.cornISSN1009—3079(print)ISSN2219—2859(online)异常凝血酶尸基因克隆表达、纯化及单克隆抗体制备与鉴定沈鹏,司进,李琦雯,渠利利,李璐蓉,严晓璐,季国忠prothrombin(DCP),avaluableserummarkerforhepatocellularcarcinoma(HCC),andtopre—parethemonoclonalantibody(mAb)againstD
3、CP.METHODS:TheDCPgenewasclonedintotheprokaryoticexpressionvectorpColdII.The薰黧;萋孵in,霎妻萎一课recombinantconstructwasexpressedinBL21Escherichiacolistrain,andtherecombinantpro.麓善?曩:由季墓凳提供。数据分析由沈鹏完成文写作由teinwithanN.terminal6..Histagwaspurifiedbynickelchelate.nitri1otriacetic.acid
4、fNi—NTA1affinitychromatography.ThepurifiedproteinwasusedtoimmunizetheBalb/cmiceformAbpreparation.ThesensitivityandspecificityofthemAbwereassessedbyindirectenzyme—linkedimmunosorbentassay(ELISA),Westernblotandimmunohistochemistry.Humandes-~-carboxyRESULTS:TherecombinantDC
5、Pproteinwasprothrombin:Cloning,successfullyexpressedandpurified,anditsmo—expressionandmonoclonallecularweightwasapproximately23kDa.Oneantibodypreparationhvbridomacelllinestablysecretinganti—DCPmonoclonalantibodynamed3B5wasobtained.Westernblotanalysisandimmunohistochemis.
6、PengShenfJinSilQi-WenLifLi—LiQufLu—RongLiIXiao’LuYan,Guo-ZhongJitryindicatedthattheDCPmAbshowedspecificcombinationwithDCP.ELISAdetectionindiPengShenfLu。RongLilXiao。LuYahlGuo。ZhongJiIcatedthatthetiterofthemAbwas1:2.4310.InstituteofDigestiveEndoscopyandMedicalCenterforThei
7、mmunoglobulinisotypeofthemAbwasDigestiveDiseases,theSecondAfiliatedHospitalofNan—identifiedasIgG2a.jingMedicalUniversity,Nanjing210011,JiangsuProvince,ChinaJinSi,Qi—WenLi,Li-LiQu,DepartmentofLaboratoryCONCLUSION:ThesuccessfulpreparationofDCPMedicine.theSecondA伍liatedHosp
8、italofNanjingMedi—mAbmaylayasolidfoundationfortheresearchofcalUniversity,Nanjing21O0l1.JiangsuProvince.ChinaSupportedby:theScienceandTechnologyDevelopmenttheuseofDCPinearlydiagnosisofHCC.ProjectofNanjing.No.2Ol104029;theMedicalKevTlalentFoundationofJiangsuProvince.No.RC2
9、0l1081:andthe⑥2013BaishidengPublishingGroupCo.,Limited.AIMedicalScienceandTechnologyDevelopmentProgramo
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