Development of rapid one-step immunochromatographic assay
Development of rapid one-step immunochromatographic assay
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DOI：
10.1006/meth.2000.1036
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发表时间：
2000-09-01
期刊：
影响因子：
--
通讯作者：
Kim, YS
中科院分区：
文献类型：
--
作者：
Paek, SH;Lee, SH;Kim, YS
利用免疫层析的概念，建立了一步免疫分析系统。该系统使用了两种不同的抗体来结合分析物分子的不同表位：一种抗体用信号发生器(例如胶体金)标记，将其置于玻璃纤维膜上预定位置的干燥状态，另一种抗体固定在硝酸纤维素膜的表面。三种膜，一种带有示踪剂，一种具有固定化抗体，以及一种纤维素膜作为介质锡的吸收剂(从下到下的顺序)，被附着在塑料薄膜上并切割成条。从免疫条底端吸收的含有分析物的水介质溶解标记的抗体，形成的抗原-抗体结合络合物通过毛细作用产生的流动输送到下一层硝酸纤维素膜。该复合体随后与固定化抗体反应，产生与分析物浓度成比例的信号。免疫反应物向结合配对的对流传质使得无需处理试剂即可进行分析。然而，该反应是在非平衡条件下进行的，这导致与在平衡模式下进行的分析(例如，ELISA)相比，灵敏度降低。为了最大限度地减少这种牺牲，确定了控制系统性能的主要因素，然后在最佳条件下设计了系统。(C)2000年学术出版社。
An analytical system for a one-step immunoassay has been constructed using the concept of immunochromatography. The system employed two different antibodies that bound distinct epitopes of an analyte molecule: an antibody labeled with a signal generator (e.g., colloidal gold), which was placed in the dry state at a predetermined site on a glass-fiber membrane, and another antibody immobilized on the surface of a nitrocellulose membrane. Three membranes, one with the tracer, one with immobilized antibody, and a cellulose membrane as the absorbent of medium tin a sequence from the bottom), were attached to a plastic film and cut into strips. Aqueous medium containing analyte absorbed from the bottom end of the immunostrip dissolved the labeled antibody, and the antigen-antibody binding complex formed was transported into the next nitrocellulose membrane by the flow caused by capillary action. The complex subsequently reacted with the immobilized antibody, which generated a signal in proportion to the analyte concentration. The convective mass transfer of the immunoreactant to the binding partner allowed the assay to be performed with no handling of reagents. The reaction, however, was carried out under nonequilibrium conditions, which resulted in decreased sensitivity as compared with assays performed in an equilibrium mode (e.g., ELISA). To minimize such sacrifice, major factors that control system performance were identified and the system was then devised under optimal conditions. (C) 2000 Academic Press.