Jeney C, Dobay O, Lengyel A, Adám E, Nász I
Institute of Microbiology, Semmelweis Medical School, Budapest, Hungary.
J Immunol Methods. 1999 Mar 4;223(2):137-46. doi: 10.1016/s0022-1759(98)00185-9.
We propose a new method in the field of ELISA optimization using an experimental design called the Taguchi method. This can be used to compare the net effects of different conditions which can be both qualitative and quantitative in nature. The method reduces the effects of the interactions of the optimized variables making it possible to access the optimum conditions even in cases where there are large interactions between the variables of the assay. Furthermore, the proposed special assignment of factors makes it possible to calculate the biochemical parameters of the ELISA procedure carried out under optimum conditions. Thus, the calibration curve, the sensitivity of the optimum assay, the intra-assay and inter-assay variability can be estimated. The method is fast, accessing the results in one step, compared to the traditional, time-consuming 'one-step-at-a-time' method. We exemplify the procedure with a method to optimize the detection of ScFv (single chain fragment of variable) phages by ELISA. All the necessary calculations can be carried out by a spreadsheet program without any special statistical knowledge.
我们提出了一种在酶联免疫吸附测定（ELISA）优化领域使用田口方法（一种实验设计）的新方法。该方法可用于比较不同条件的净效应，这些条件在性质上可以是定性的和定量的。该方法减少了优化变量之间相互作用的影响，使得即使在测定变量之间存在较大相互作用的情况下也能够确定最佳条件。此外，所提出的特殊因子分配使得计算在最佳条件下进行的ELISA程序的生化参数成为可能。因此，可以估计校准曲线、最佳测定的灵敏度、批内和批间变异性。与传统的耗时的“一次一步”方法相比，该方法速度快，一步即可获得结果。我们以一种优化通过ELISA检测单链可变区片段（ScFv）噬菌体的方法为例来说明该程序。所有必要的计算都可以通过电子表格程序进行，无需任何特殊的统计知识。
