Jensenius J C, Siersted H C, Johnstone A P
J Immunol Methods. 1983;56(1):19-32. doi: 10.1016/0022-1759(83)90045-5.
Simple, reliable semiautomatic radioimmunoassays have been developed for the measurement of human immunoglobulins (IgG, IgA, IgM, IgK and Ig lambda). The assays are based on the separation of free from antibody-bound radiolabelled fragments of immunoglobulin by precipitation with 13% polyethylene glycol. The precipitate is harvested and washed on glass fibre filters by means of a commercial cell harvester. Radiolabelled lambda chain from Bence Jones urine was used for lambda chain assay and Fab' gamma from pooled IgG for the kappa assay. Labelled Fab or Fc fragments of IgG, IgA and IgM were used for the class specific assays. Selected commercial antisera were used throughout. The sensitivity limits of the assays performed according to the standard procedure were about 20 ng Ig per ml. The assays have been used for the quantification of circulating immune complexes and both cellular and secreted immunoglobulin produced in cell cultures. The results demonstrate the importance of the simultaneous quantification by L chain and H chain specific assays.
已开发出简单、可靠的半自动放射免疫分析法,用于检测人免疫球蛋白(IgG、IgA、IgM、IgK和Igλ)。这些分析方法基于用13%聚乙二醇沉淀,将免疫球蛋白的游离放射性标记片段与抗体结合的片段分离。沉淀物通过商用细胞收获器收集并在玻璃纤维滤器上洗涤。来自本斯·琼斯尿液的放射性标记λ链用于λ链分析,来自混合IgG的Fab'γ用于κ链分析。IgG、IgA和IgM的标记Fab或Fc片段用于类别特异性分析。全程使用选定的商用抗血清。按照标准程序进行的分析的灵敏度极限约为每毫升20 ng Ig。这些分析方法已用于定量循环免疫复合物以及细胞培养物中产生的细胞免疫球蛋白和分泌免疫球蛋白。结果表明通过轻链和重链特异性分析同时定量的重要性。