重组抗人 CD22嵌合抗体 SM03的 125 I标记及生物活性
Bioactivity assays and application of 125I radiolabeled human mouse chimeric anti-CD22 monoclonal antibody, SM03
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摘要: 摘要: 目的 探讨采用125I标记的重组抗人CD22嵌合抗体SM03的生物活性测定以及应用。方法 将单克隆抗体SM03用Iodogen法标记，用Sephacryl S-300 HR色谱主分离标记混合物，测定分离后样品的纯度与浓度。采用竞争结合法和酶联免疫吸附法（ELISA)测定纯化后样品的活性。最后用标记后的SM03筛选CD22抗原表达量最多的细胞。结果 抗体经过放射性标记后纯度>99%,回收率>67%。125I-SM03的生物活性与未标记抗体没有显著性差异。在Raji,Daudi和Ramos细胞中，SM03与Ramos细胞的特异性结合量最多，结合量为1.3 pmol/细胞。结论 Indogen法可以用于单克隆抗体SM03的标记，标记后不会影响抗体的活性。证明了标记后的SM03具有应用价值。Abstract: Abstract: AIM To investigate the bioactivity and application of 125I radiolabed human mouse chimeric monoclonal SM03. METHODS: SM03 was radiolabeled with Indogen method. The radiolabled mixture was purified by Sephacryl S-300 HR separation chromospectry. We determined the purity and concentration of separated fraction. The radiolabeled SM03 was applied to screen which cell line express maximum CD22 antigen. RESULTS: The purity and recovery of radiolabeled SM03 were >99% and >47%, respectively. The bioactivity of 125I labeled SM03 and non-labeled SM03 did not have significant difference in statistics. Ramos cell line had the strongest special radioactivity when bound with SM03 in Raji, Daudi and Ramos cell lines. CONCLUSION: Human mouse chimeric anti-CD22 monoclonal antibody can be radiolabeled with Indogen method. The radiolabel will not affect the activity of SM03.
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