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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。
Objective To develop recombinant human bone sialoprotein (rhBSP) monoclonal antibody (mAb) and identify its characteristics. Methods Balb / c mice were immunized with purified rhBSP, and the anti-rhBSP mAb was prepared by hybridoma technique. The IgG subclasses were identified by subtype identification reagents and the specificity and titer of mAbs were identified by ELISA. Results Two anti-rhBSP hybridoma cell lines, AHB1 and AHB5, which can stably secrete specific mAbs were obtained. The subclasses of IgB were IgG2a and IgG1, and the light chains were all κ. The titer was 1 × 10-3 and 1 × 10-7. Ascites mAb purification by Protein A affinity column, the purity of 92% or more. Conclusion Obtaining anti-rhBSP mAb is the basis for further study on the biological function of BSP and its application in clinical diagnosis.