抗死亡受体-5单克隆抗体的制备及特性鉴定  

Preparation and characterization of mon-oclonal antibody against DR5

作　　者：白慧玲[1] 赵粤萍[2] 刘广超[2] 马远方[2] 

机构地区：[1]河南大学免疫学研究所细胞与分子免疫学实验室,河南开封475001 [2]河南大学细胞与分子免疫学实验室,河南开封475001

出　　处：《细胞与分子免疫学杂志》2006年第4期514-516,520,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：河南省杰出人才创新基金(0321001800);河南省医学科技创新人才基金资助(2002-119)

摘　　要：目的：研制抗DR5单克隆抗体（mAb），并鉴定其特性。方法：以纯化的DR5免疫BALB／c小鼠，采用杂交瘤技术制备抗DR5mAb。用Ig亚类ELISA试剂盒鉴定抗DR5mAb的亚类。用ELISA法测定sDR5对抗DR5 mAb与DR5结合的阻断作用及流式细胞术检测与Jurkat细胞膜上DR5结合的阻断作用。以鉴定mAb的特异性。用间接ELISA法检测腹水mAb的效价、亲和常数并进行表位分析。结果：获得4株可分泌抗DR5 mAb的的杂交瘤细胞系YM366EC、YM366ED、YM369F5和YM369E6。4株mAb的Ig亚类均为IgG1；腹水mAb效价为1×10^-4-5×10^-6；亲和常数为1×10^9水平，4株mAb可识别2种不同的抗原表位。结论：获得4株抗DR5的mAb，为进一步用于临床诊断和实验研究创造了条件。AIM： To prepare monoclonal antibodies （mAb） against DR5 and characterize their properties. METHODS： BALB/c mice were immunized with DRS, and then mAb was prepared by hybridoma technique. Ig subclass and specificity of mAbs was analyzed by ELISA and flow cytometry, respectively. The titres of mAbs in ascitic fluid, relative affinity and epitopes recognized by mAbs were determined by indirect ELISA. RESULTS： Four hybridoma cell lines secreting anti DR5 mAbs were obtained. Their Ig subclass belonged to IgG1. The titers of 4 mAbs in ascitic fluid were 1 × 10^ -4 -5 × 10^ -6. Affinity constant of mAbs were 1 × 10^9. They recognized 2 different epitopes on DR5 molecule. CONCLUSION： Four mAbs against DR5 are prepared successfully, which provides useful reagent for clinical diagnosis and further research.

关 键 词：DR5 单克隆抗体 特性鉴定 亲和常数 

分 类 号：R392.11[医药卫生—免疫学]