Wang R, Merrill B, Maggio E T
Clin Chim Acta. 1980 Mar 28;102(2-3):169-77. doi: 10.1016/0009-8981(80)90030-3.
A simple 2-step solid-phase fluoroimmunoassay for the determination of serum immunoglobulins is described. The method employs a stabilized solid-phase immunoadsorbent consisting of antigen immobilized on a cellulose acetate/nitrate disc attached to a plastic StiQTM sampler. The polymeric disc serves both as a substrate for immobilization and as a highly uniform surface upon which very precise fluorescence determinations may be made. The assays rely upon the reaction of a precise and limiting amount of mono-specific fluorescent antibody with the specific antigen present in a test sample. Residual unreacted fluorescently labeled antibody is allowed to bind to the immunoadsorbent; non-specifically bound antibody is removed in a wash step. The amount of labeled antibody bound is inversely proportional to the amount of antigen present in the test sample. The fluorescence of the bound labeled antibody is measured using a FIAX surface-reading fluorometer. Comparison of this new procedure with the commercially available (4-step) FIAX assay, which requires generation of immunoadsorbent in situ, demonstrated excellent correlation between the two methods. The new 2-step procedure provides results in a much shorter period of time and requires one-half to one-eighth of the operator time required for the other available solid-phase fluoroimmunoassays.
本文描述了一种用于测定血清免疫球蛋白的简单两步固相荧光免疫测定法。该方法采用一种稳定的固相免疫吸附剂，它由固定在附着于塑料StiQTM采样器的醋酸纤维素/硝酸纤维素圆盘上的抗原组成。聚合物圆盘既作为固定的底物，又作为一个高度均匀的表面，在其上可以进行非常精确的荧光测定。该测定法依赖于精确且限量的单特异性荧光抗体与测试样品中存在的特异性抗原的反应。使残留的未反应荧光标记抗体与免疫吸附剂结合；在洗涤步骤中去除非特异性结合的抗体。结合的标记抗体的量与测试样品中存在的抗原量成反比。使用FIAX表面读数荧光计测量结合的标记抗体的荧光。将该新方法与市售的（四步）FIAX测定法进行比较，后者需要原位生成免疫吸附剂，结果表明这两种方法具有极好的相关性。新的两步法在更短的时间内提供结果，并且所需的操作人员时间仅为其他可用固相荧光免疫测定法的二分之一至八分之一。
