抗髓鞘少突胶质细胞糖蛋白抗体放射免疫测定方法的建立 |
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引用本文: | 叶好好,厉向,夏君慧,张琦,张旭.抗髓鞘少突胶质细胞糖蛋白抗体放射免疫测定方法的建立[J].浙江医学,2011,33(7):989-992. |
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作者姓名: | 叶好好 厉向 夏君慧 张琦 张旭 |
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作者单位: | 1. 温州医学院附属第一医院神经内科,325000 2. 温州医学院核医学教研室 |
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摘 要: | 目的 建立以125I -抗髓鞘少突胶质细胞糖蛋白(MOG)一硫氧还融合蛋白(MOG融合蛋白)介导的放射免疫分析技术(RIA)作为测定实验性自身免疫性脑脊髓炎(EAE)小鼠血清抗MOG抗体的新方法.方法 Na125I标记MOG融合蛋白,以MOG融合蛋白免疫C57BL/6小鼠建立EAE模型,将125I -MOG融合蛋白与小鼠血清标本结合,孵育后与兔抗鼠IgG结合,收集沉淀,用γ放射性检测仪测定抗MOG抗体水平.结果 成功制备纯度较高的125I-MOG融合蛋白,当实验条件为Na125I 投料量1mCi,MOG融合蛋白量60μg时,获得较好的标记率48.3631%,最高的比放射性320 043.7 Bq/μg.应用新型放射免疫法测定小鼠血清抗MOG抗体,MOG组血清抗MOG抗体水平与其他组比较差异有统计学意义(P<0.01).结论 成功建立了一种较为敏感的测定血清抗MOG抗体的放射免疫方法.
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关 键 词: | 髓鞘少突胶质细胞糖蛋白自身抗体 多发性硬化 放射免疫分析 |
Establishment of radioimmunoassay for detecting myelin oligodendrocyte glycoprotein autoantibody |
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Institution: | YE Haohao,LI Xiang,XIA Junhui, et al. Department of Neurology, the First Affiliated Hospital of Wenzhou Medical College, Wenzhou 325000, China |
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Abstract: | Objective To establish a radioimmunoassay(RIA) technique for detecting anti-myelin oligodendrocyte glycoprotein autoantibodies(anti-MOG antibody). Methods MOG fusion protein was labeled by Na125^I We had established exprimental autoimmune encephalomyelitis (EAE) mice immunized by MOG fusion protein. Serum from mice binding with ^125I -MOG fusion protein was incubated overnight; and rabbit anti-mouse IgG was added as second antibody. Finally the precipitation was collected and the level of anti- MOG antibody was measured. Results The ^125I -MOG fusion protein with high purity was produced. When MOG fusion protein (60μg) was radioiodinated with lmCi Na^125I, the radiolabeled yield was 48.3631%, and the specific activity is 320043.7 Bq / μg. Detected by the new RIA method, the level of Anti-MOG antibodies in EAE group was much higher than other groups (P〈0.01). Conclusion We have successfully established a new radioimmunoassay method for detection of anti-MOG antibody in serum of EAE mice. |
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Keywords: | Anti-myelin oligodendrocyte glycoprotein antibody Multiple sclerosis Radioimmunoassay |
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