IL1RAP单克隆抗体制备与鉴定

《IL1RAP单克隆抗体制备与鉴定》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、·962·中国实验血液学杂志JournalofExperimentalHematology2015;23(4):962－965文章编号(ArticleID):1009－2137(2015)04－0962－04·论著·IL1ＲAP单克隆抗体的制备与鉴定1111，2*1，2*尹玲玲，阮素红，田宇，徐开林，赵恺12徐州医学院血液病研究所，骨髓干细胞重点实验室;徐州医学院附属医院血液科，江苏徐州221002摘要目的:制备针对白血病干细胞(LSC)表面标志蛋白IL1ＲAP(IL-1receptoraccessorypro

2、tein)的特异性单克隆抗体并对其进行鉴定。方法:杂交瘤细胞3H6E10、10D8A7种植BALB/c小鼠腹腔，收获腹水，纯化后得到特异性抗人IL1ＲAP的单克隆抗体。分别应用非变性聚丙烯酰胺凝胶电泳(Nondenaturing-PAGE)、ELISA和Westernblot法对纯化单克隆抗体分别进行抗体纯度、效价和敏感性检测。结果:获得了两种IL1ＲAP纯化单克隆抗体，命名为3H6E10McAb和10D8A7McAb，其纯度分别为95%和94%;两种纯化的单克隆抗体效价为1∶81000，且能特异性识别IL1Ｒ

3、AP纯化蛋白、正常人及白血病病人的细胞内源性蛋白。结论:本研究成功制备了能特异性结合人IL1ＲAP的纯化单克隆抗体，为将来有效清除体内LSC提供了新途径。关键词IL1ＲAP;单克隆抗体;白血病干细胞中图分类号Ｒ733．7文献标识码Adoi:10．7534/j．issn．1009-2137．2015．04．010PreparationandAssessmentofIL1ＲAPMonoclonalAntibody1111，2*1，2*YINLing-ling，ＲUANSu-Hong，TIANYu，XUKaiLin，

4、ZHAOKai12BloodDiseasesInstitute，KeyLabatoryofBoneMarrowStemCells，XuzhouMedicalCollege，DepartmentofHematology，TheAffilia-tedHospitalofXuzhouMedicalCollege，Xuzhou221002，JiangsuProvince，China*Co-correspondingAuthor:XUKai-Lin，SeniorPhysician．E-mail:limd@163．com;

5、ZHAOKai，AssociateSeniorScientist．E-mail:kainyzhao@163．comAbstractObjective:ToprepareandidentifyhumanmonoclonalantibodyagainstIL-1receptoraccessoryprotein(IL1ＲAP)，whichisanewidentifiedsurfacemarkerforleukemiastemcells(LSC)．Methods:BALB/cmicewereinoculatedintr

6、aperitoneallywithhybridomacells(3H6E10，10D8A7)andtheirasciteswerecollected．Themonoclonalantibodyagainsthu-IL1ＲAPspecificallywaspurifiedfromascites，thenondenaturing-PAGE，ELISAandWesternblotwereusedtodetectthepurity，titerandsensitivityofantibody．Ｒesults:Twopur

7、ifiedantibodieswereobtainedandnamedas3H6E10McAband10D8A7McAb，whosepuritywas95%and94%respectively．Thetiteroftwopurifiedmonoclonalantibodieswas1∶81000andspecificconjugationofIL1ＲAPpurifiedproteinandendogenousproteinfromnormalpeopleandleukemiapatientswithpurifi

8、edantibodieswereconfirmed．Conclusion:Thepurifiedmonoclonalantibodieswhichcanspecificallybindtohu-IL1ＲAParesuccessfullyprepared，thusprovidingnovelwaytoeffectivelyclearLSCinthefuture．KeywordsIL1ＲA