Preparation, characterization, and activity analysis of monoclonal antibodies against human death receotor 5
YANG Donghai
ZHUANG Guohong
LIU Zhongchen
ZHANG Changgong
HE Qiong
LI Wenzhu
Abstract：Objective To prepare monoclonal antibodies(mAbs)against DR5,and characterize their biological properties by using ELISA,flow cytometry,and SDS-PAGE.Methods Balb/c mice were immunized with soluble DR5(sDR3),and then mAbs was prepared by hybridoma technique.Ig subclass and specificity of mAbs were analyzed by ELISA and SDS-PAGE,respectively.The titres of mAbs in ascitic fluid were determined by indirect ELISA.Flow cytometry was utilized to detect the tumor cell apoptosis-indu-cing activity of the mAbs and the DR3 expression on the tumor cell surface.Results One hybridoma cell line secreting anti-DR5 mAbs were obtained,named R150.DR5 mAb R150 Ig subclass belonged to IgG.whose titer in ascitic fluid was 1×106.The expression of DR5 on different tumor cell SHrfaces could be Sensitively detected by flow cytometry.Conclusion Anti-DR5-mAbs against DR5 have been prepared successfully,which possesse high titers and specificity and cpuld be used in membrane DR5 and sDR5 testing.
Keywords：death receptor 5TRAILmonoclonal antibodyapoptosis
Publication Date：2009-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 671-675 )