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时间:2020-04-22
《抗胰腺癌干细胞单克隆抗体的筛选和鉴定-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、·1492·孙立新,等抗胰腺癌干细胞单克隆抗体的筛选和鉴定抗胰腺癌干细胞单克隆抗体的筛选和鉴定孙立新,遇珑,解亦斌,莫文秀,刘辉琦,杨治华,冉宇靓,孙力超ScreenandidentificationofmonoclonalantibodyagainstpancreaticcancerstemcellsSunLixin,YuLong,XieYibin,MoWenxiu,LiuHuiqi,YangZhihua,RanYuliang,SunLichaoStateKeyLaboratoryofMolecularOncology,NationalEngineerin
2、gLaboratoryforHigh—levelExpressioninMammalianCells,Zhong-guaneunScienceP口OpenLaboratoryofEngineeringAntibodyDrugsandCancerBiomarker,CancerInstitute~Hospital,ChineseAcademyofMedicalSciences(CAMS),PekingUnionMedicalCollege,Beijing100021,China;eDepartmentofAbdominalSurgicalOneologyCan
3、cerHospital,ChineseAcademyofMedicalSciences(CAMS),Beng100021,China.【Abstract】0bjective:Topreparefunctionalmonoclonalantibodies(mcAbs)againstpancreaticcancerstemcell,soastoprovidecandidateantibodydrugsforstemcell—targetedtherapyofpancreaticcancer.Methods:HPACcellswerecuhuredinserum—
4、freemedium.FlowcytometrywasusedtodetecttheexpressionofCD133inHPACcellsandspherecells.TheefectsofhybridomamcAbsonself—renewal,proliferationandchemosensitivitytoGemcitabineofHPACparentorspherecellswereidentifiedbyserum—freesuspensioncultureandCCK一8assay.Results:HPACcellscouldsurvive.
5、proliferateandformspherecellsinserum—freemedium.Thesphere—formingratewas4.8%±0.6%.ThepercentageofCD133cellspopulationinspherecellsincreasedby11.6foldscomparedtoHPACcells.ThepositiverateofCD133inHPACspherecellswassignificantlyhigherthanthatinHPACparentcells.Amongthe20candidatemcAbs.
6、3mcAbscanrecognizeCD133cellsinHPACspherecells.Thepositiverateof15E9was3.5%.FurtherinvestigationshowedthatmcAbs15E9significantlysuppressedthesphereformationabilityofHPACcells,withtheinhibitoryratesbeing30.4%.TheIC50was30.8nmol/Lin15E9+Gemcitabinegroup,and58.1nmoL/LinmIgM+Gemcitabine
7、group.Conclusion:WehavesuccessfullyscreenedandidentifiedmcAbs15E9fromlargemcAblibraryagainstpancreaticcancerstemcell,whichcanspecificallyrecognizeCD133HPACcellsandinhibitself—renewa1.mcAb15E9couldbethecandidateantibodydrugtargetingcancerstemcel1.【Keywords】pancreaticcancer;cancerste
8、mcell;monoelonalantibody;t
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