Shamsuddin A M, Harris C C
Arch Pathol Lab Med. 1983 Oct;107(10):514-7.
Using the characteristic of a high-affinity complex between enzyme-conjugated avidin and biotin, we improved the techniques of enzyme-linked immunosorbent assay and ultrasensitive enzymatic radioimmunoassay. The method included biotinylation of a specific antibody, complexing of a biotinylated antibody with avidin-alkaline phosphatase conjugate, and addition of the antibody-conjugate complex (with or without a standard amount of antigen for competitive or noncompetitive assays, respectively) to the microtiter wells containing antigen in the solid phase. Once biotinylated, antibodies could be stored for several months. Complexing of biotinylated antibody with avidin-alkaline phosphatase conjugate could also be performed once, and the stock solution could be used for several days. Thus, the steps of enzyme immunoassays were reduced to the addition of only antibody-conjugate complex and enzyme substrate. The improved assays, biotin-avidin-linked enzyme immunoassay and biotin-avidin-linked ultrasensitive radioimmunoassay, were simpler, quicker, less expensive, and more sensitive than their standard counterparts.
利用酶联抗生物素蛋白与生物素之间高亲和力复合物的特性，我们改进了酶联免疫吸附测定法和超灵敏酶免疫放射测定法。该方法包括特异性抗体的生物素化、生物素化抗体与抗生物素蛋白 - 碱性磷酸酶偶联物的复合，以及将抗体 - 偶联物复合物（分别用于竞争性或非竞争性测定，有或没有标准量的抗原）加入到含有固相抗原的微量滴定孔中。一旦进行生物素化，抗体可以储存数月。生物素化抗体与抗生物素蛋白 - 碱性磷酸酶偶联物的复合也可以一次性完成，储备溶液可以使用数天。因此，酶免疫测定的步骤减少到仅添加抗体 - 偶联物复合物和酶底物。改进后的测定方法，即生物素 - 抗生物素蛋白连接酶免疫测定法和生物素 - 抗生物素蛋白连接超灵敏放射免疫测定法，比其标准对应方法更简单、更快、更便宜且更灵敏。
