P2X7受体短肽单克隆抗体的制备及鉴定.pdfVIP

生物技术通报 ·研究报告· BIOTECHNOLOGYBULLETlN 2012：￡lZ第12期 P2X7受体短肽单克隆抗体的制备及鉴定 谭超1’2韩莉2神谢静2 (1三峡大学医学院，宜昌443002；2三峡大学第一临床医学院宜昌市中心人民医院检验科，宜昌443002) 摘要： 旨在制备与鉴定鼠抗P2X7受体的蛋白单克隆抗体。以人P2X7受体的胞外段制备短肽作为抗原，皮下注射免疫 Balb／c小鼠。分离小鼠脾脏B淋巴细胞与骨髓瘤细胞融合并培养，挑选阳性杂交瘤细胞，扩大培养，制备和鉴定P2X7其生物学效 应。结果显示，获得1株稳定分泌抗人P2X7受体的单克隆抗体的杂交瘤细胞株，所分泌的单抗类型重链为IgGl，轻链为1c；该株 X 杂交瘤细胞腹水效价为1：6．4104；传30代及液氮中保存6个月，抗体效价稳定；Western blotting检测证明该单抗与人细胞表面 体药物的开发应用、疾病的辅助诊断奠定了基础。 关键词：P2X7受体短肽单克隆抗体 The andIdentificationofP2X7Short Preparation PeptideReceptor Monoclonal Antibody TanCha01·2HanLi2Shen Xiejin92 MedicalScience Three Medicine Central (1 CollegeofChinaGorgesUniversity，Yichang443002；2DepartmentofInternalYichang People’s Medicine．theFirstClinicalMedical Three 443002) ofChina University，Yichang HospitalLaboratory College Gorges of methodwastousetheextracellular Abstract：Theaimwasto and themonoclonalantibodiestheP2X7 prepareidentify receptor．Our ofhuman to theshort asall toimmunizemouseforthreetimes．ThenB fragment beings’P2X7receptorproduce peptideantigen lymphocytes wereisolatedfromthemouse thenmixedwith cellsandcuhured．The cloneswereselectedand spleen，and myeloma positivehybridoma widely ofP2X7．The wascollectedand G column． culturedto and th