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制备重组人骨唾液酸蛋白(rhBSP)单克隆抗体(mAb)并对其性质进行鉴定。采用杂交瘤技术制备mAb,并测定亚型、效价、亲和常数、抗原表位,同时应用竞争抑制ELISA法分析其特异性。结果:得到2株杂交瘤阳性细胞株AHB2和AHB6,亚型为IgM和IgG1,轻链均为κ型;AHB2和AHB6的效价分别为10-4,10-6,亲和常数分别约为3·01×108M-1,2·94×1010M-1。检测发现,AHB2和AHB6均与大鼠血清有弱阳性反应。通过竞争抑制ELISA法分析得出AHB2和AHB6两株抗体针对BSP的相同抗原表位。采用Envision法进行BSP免疫组化的检测研究。成功制备了rhBSP的特异性mAb,为进一步研究BSP的生物学功能和临床诊断实验研究打下基础。
Preparation of recombinant human bone sialic acid (rhBSP) monoclonal antibody (mAb) and its nature were identified. MAb was prepared by hybridoma technology, and the subtype, titer, affinity constant and antigen epitope were determined. At the same time, its specificity was analyzed by competitive inhibition ELISA. Results: Two hybridoma positive cell lines, AHB2 and AHB6, were obtained. The subtypes were IgM and IgG1, and the light chains were all κ. The titer of AHB2 and AHB6 were 10-4 and 10-6, respectively, and the affinity constants were about 3 · 01 × 108M-1,2 · 94 × 1010M-1. Test found that, AHB2 and AHB6 are weak positive reaction with rat serum. By competition inhibition ELISA analysis AHB2 and AHB6 two antibodies against BSP the same epitope. Detection of BSP Immunohistochemistry with Envision Method. The specific mAb of rhBSP was successfully prepared, which laid the foundation for further study on the biological function and clinical diagnosis of BSP.