肿瘤M2型-丙酮酸激酶单克隆抗体的制备及免疫放射分析方法的建立

Preparation of the monoclonal antibodies against tumor M2-type pyruvate kinase and establishment of immunoradiometric analysis for detection

    摘要
  • 摘要:
    目的 制备肿瘤M2型-丙酮酸激酶(M2-PK)单克隆抗体，并建立免疫放射分析的检测方法。
    方法 用肿瘤M2-PK抗原免疫小鼠，经细胞融合及筛选后得到一对单克隆抗体，建立免疫放射分析的方法，并评价该检测方法的灵敏度、特异度及精密度。
    结果 获得了1A6和3F8两株效价高、特异度好的单克隆抗体，建立了双抗体夹心免疫放射分析方法。该检测方法的分析灵敏度为2 U/ml，临床灵敏度和特异度分别为70.5%和89.0%，提供的M2-PK的正常参考值范围是 < 14.8 U/ml，批内不精密度 < 5%，批间不精密度 < 10%。
    结论 建立的双抗体夹心免疫放射分析法可用于体检筛查及肿瘤患者的早期诊断，做到尽早治疗，提高患者的生存质量。

     

    Abstract:
    Objective To produce monoclonal antibodies against tumor M2-type pyruvate kinase(M2-PK), and to establish immunoradiometric analysis for detection.
    Methods The BALB/c mice were immunized with M2-PK antigen. Monoclonal antibodies were generated after cell fusion and screening. Established detection method of immunoradiometric analysis and then evaluated its sensitivity, specificity and coefficient of variation.
    Results Two monoclonal antibodies(1A6 and 3F8)with high-titer and high-specificity were generated and detection method of immunoradiometric analysis was established. The analytical sensitivity of immunoradiometric analysis was 2 U/ml, while its clinical sensitivity and specificity was 70.5% and 89.0% respectively. The reference value of M2-PK level was less than 14.8 U/ml according to immunoradiometric analysis. Intra-assay coefficient of variation was less than 5%, while inter-assay coefficient of variation was less than 10%.
    Conclusion This sensitive immunoradiometric analysis can be used in early diagnosis of tumor and physical examination.

     

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