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为了适用于特定抗原基质球(DASS)系统,用标记过氧化物酶的免疫球蛋白代替标记荧光素的免疫球蛋白,以了解反应的特异性、敏感性和重演性,作者进行了如下工作。抗原制备:按作者(1974)报道的方法。过氧化物酶结合免疫球蛋白的制备:按Avrameas等的两步法使猪抗人IgG(SwAHu)与辣根过氧化物酶(HRP)结合。经葡聚糖凝胶G200柱层析将结合物与游离的HRP分开。结合物的蛋白质浓度为每毫升0.01M、pH 7.3的磷酸缓冲盐水(PBS)中含0.5毫
To adapt to a specific antigen matrix sphere (DASS) system, immunoglobulins labeled with peroxidase were used instead of fluorescein labeled immunoglobulins to understand the specificity, sensitivity and repeatability of the reaction. The authors performed the following work. Antigen preparation: The method reported by the author (1974). Preparation of peroxidase-conjugated immunoglobulin: Swine anti-human IgG (SwAHu) was conjugated to horseradish peroxidase (HRP) according to the two-step procedure of Avrameas et al. Conjugates were separated from free HRP by Sephadex G200 column chromatography. The conjugate had a protein concentration of 0.5 milliliters per milliliter of phosphate buffered saline (PBS) at pH 7.3