Peterman J H, Butler J E
University of Iowa.
Biotechniques. 1989 Jun;7(6):608-15.
Solid-phase immunoassays such as the ELISA are in routine use in many areas of biological research. Data from these assays are analyzed in a variety of ways, frequently without taking into account the immunochemical principles of the assay. The Reference Standard Method is often used and is suitable and convenient for obtaining concentration (or activity) values from the antigen-specific ELISA or spRIA, sandwich assays, and inhibition assays. The standard curve required for this method may be obtained by simple linear regression analysis of logarithmic or logitlogarithmic transformed data obtained from titration of the reference standard. The shape of the logarithmic plot of the reference standard provides information on the performance of the assay. Examining data from multiple dilutions of the samples is essential to assure that each titrates with the same slope as does the reference standard; the analysis routine must permit this comparison to be made. ELISANALYSIS is a program for the IBM PC which was developed to perform such analyses. It is presented here as a model, with sufficient information provided for the development of similar analytical routines by interested users. This approach to ELISA data analysis is presented as an alternative to complicated empirical curve-fitting systems and simple endpoint methods, which can be immunochemically misleading or, in some cases, even invalid. The consistent use of the described routines would encourage greater uniformity in the means of data interpretation and thereby enhance our understanding of immunobiology.
诸如酶联免疫吸附测定（ELISA）之类的固相免疫测定法在生物学研究的许多领域中都有常规应用。这些测定法所获得的数据有多种分析方式，而常常没有考虑到测定法的免疫化学原理。参考标准方法经常被使用，并且适用于从抗原特异性ELISA或spRIA、夹心测定法和抑制测定法中获取浓度（或活性）值，既合适又方便。此方法所需的标准曲线可通过对参考标准滴定所得的对数或对数-对数转换数据进行简单线性回归分析来获得。参考标准的对数图形状可提供有关测定法性能的信息。检查样品多个稀释度的数据对于确保每个样品的滴定斜率与参考标准相同至关重要；分析程序必须允许进行这种比较。ELISANALYSIS是一个为IBM个人计算机开发的程序，用于执行此类分析。在此将其作为一个模型呈现，为感兴趣的用户提供了足够的信息以开发类似的分析程序。这种ELISA数据分析方法是作为复杂经验曲线拟合系统和简单终点方法的替代方法提出的，后两种方法在免疫化学上可能会产生误导，在某些情况下甚至是无效的。持续使用所述程序将有助于在数据解释方法上实现更大的一致性，从而增进我们对免疫生物学的理解。
