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

摘要

Objective To establish a radioimmunoassay(RIA) technique for detecting anti- myelin oligodendrocyte glycoprotein autoantibodies (anti- MOG antibody).Methods MOG fusion protein was labeled by Na125I.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 1mCiNa125I, 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.%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.