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

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.