Freytag J W, Lau H P, Wadsley J J
Clin Chem. 1984 Sep;30(9):1494-8.
We describe an affinity-column-mediated, enzyme-linked immunometric assay that is highly sensitive and adaptable to automation. Digoxin is the model test analyte. A comparison of digoxin with its analog, ouabain, for use as the immobilized ligand on the affinity column showed ouabain to be superior. We also report the effect of column elution rate. Antibody-enzyme conjugates prepared with the monovalent Fab'-fragment and the divalent F(ab')2-fragment coupled to beta-galactosidase are compared in terms of their overall assay performance. Although the monovalent Fab'--beta-galactosidase conjugate yields a more sensitive assay and dose-response curves that are linear over a wider range, the divalent F(ab')2--beta-galactosidase conjugate provides an assay with adequate sensitivity and extremely good precision, and is generally easier to synthesize reproducibly. This fully automated, rapid, and precise assay for digoxin is compatible with the Du Pont aca discrete clinical analyzer.
我们描述了一种亲和柱介导的酶联免疫分析方法，该方法高度灵敏且适用于自动化操作。地高辛是模型检测分析物。将地高辛与其类似物哇巴因作为亲和柱上的固定配体进行比较，结果显示哇巴因更具优势。我们还报告了柱洗脱速率的影响。对用单价Fab'-片段和与β-半乳糖苷酶偶联的二价F(ab')2-片段制备的抗体-酶缀合物的整体分析性能进行了比较。尽管单价Fab'-β-半乳糖苷酶缀合物产生的分析方法更灵敏，剂量-反应曲线在更宽范围内呈线性，但二价F(ab')2-β-半乳糖苷酶缀合物提供的分析方法具有足够的灵敏度和极高的精密度，并且通常更容易可重复地合成。这种用于地高辛的全自动、快速且精确的分析方法与杜邦aca离散临床分析仪兼容。
