抗重组人骨唾液酸蛋白单克隆抗体的制备及鉴定  

Preparation and characterization of monoclonal antibodies against recombinant human bone sialoprotein

作　　者：王捷[1] 刘增田[1] 杨太成[1] 杨静[1] 冼江[1] 刘大志[1] 

机构地区：[1]广州军区广州总医院医学实验中心,广东广州510010

出　　处：《中国生化药物杂志》2006年第6期323-325,共3页Chinese Journal of Biochemical Pharmaceutics

基　　金：广东省自然科学基金研究团队项目(No20023001);广东省自然科学基金自由申请项目(No04003534)

摘　　要：目的研制重组人骨唾液酸蛋白(rhBSP)单克隆抗体(mAb),并鉴定其特性。方法以纯化的rhBSP免疫Balb/c小鼠,采用杂交瘤技术制备抗rhBSP mAb;用亚型鉴定试剂条鉴定IgG亚类;ELISA鉴定mAb的特异性和效价。结果获得2株能稳定分泌特异性mAb的抗rhBSP的杂交瘤细胞系AHB1和AHB5,Ig亚类分别为IgG2a和IgG1,轻链均为κ型,其效价分别为1×10-3和1×10-7。腹水mAb经Protein A亲和色谱柱纯化后,纯度达92%以上。结论获得抗rhBSP的mAb,为进一步研究BSP的生物学功能和用于临床诊断实验研究创造了条件。Purpose To prepare monoclonal antibodies （mAb） against recombinant human bone sialoprotein （rhBSP） and to characterize their properties. Methods Balb/c mice were immunized with rhBSP, and mAb was prepared by hybridoma technique, lgG subclass was detected by mAb Isotyping Test Kit. The titres and specificity of mAbs was determined by ELISA. Results Two hybridoma cell lines secreting anti-rhBSP mAbs were obtainod and named AHBI and AHB5. Their IgG subclass belonged to IgGZa and lgGl respectively and both with κ light chains. The titers of AHBI and AHB5 in ascitic fluid were 1 ×10^-3,and 1 ×10^-7 respectively. The purity of mAb was over 92 % alter protein A affinity chromatography. Conclusion Two mAbs against rhBSP have been prepared successfully which provide useful reagent for further studying the biological function of BSP and clinical diagnosis.

关 键 词：人骨唾液酸蛋白 单克隆抗体 重组 制备 特性鉴定 

分 类 号：R392.11[医药卫生—免疫学]