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

《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。

1、'

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