MEASUREMENT OF SERUM FERRITIN BY A 2-SITE IMMUNORADIOMETRIC ASSAY
MEASUREMENT OF SERUM FERRITIN BY A 2-SITE IMMUNORADIOMETRIC ASSAY
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DOI：
10.1016/0003-2697(74)90347-9
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发表时间：
1974-01-01
影响因子：
2.9
通讯作者：
COOK, JD
中科院分区：
文献类型：
--
作者：
MILES, LEM;LIPSCHITZ, DA;COOK, JD
通过铁蛋白溶液与固相抗（人铁蛋白）反应，然后进行第二次反应，其中不溶性产物与纯化的放射性标记的抗（人铁蛋白）孵育，进行人血清铁蛋白的2-位点免疫放射测定（2-位点IRMA）。未反应的标记抗体保留在溶液中并被洗掉。随着铁蛋白量的增加，固相中的放射性增加。评价了影响测定的因素，包括（a）固相抗体的浓度和稳定性，（B）温度、反应时间和试剂浓度的变化，（c）抗体包被管的稳定性和储存，（d）管洗涤周期的影响，（e）血清蛋白和抗凝剂的影响，（f）铁蛋白的器官特异性。在高剂量下观察到剂量反应的反常下降。统计分析，剂量插值和自动数据处理进行了推广的logit/log方法和计算机程序中使用的常规放射免疫分析。方差对剂量-反应曲线上位置的依赖性与放射免疫测定系统中观察到的不同，并反映在更大的有效测定范围内。2-现场IRMA也可能在试剂稳定性、特异性、抗原保护和自动化适用性方面具有优势。2-位点IRMA的性质与通常的IRMA测定系统密切相关，但2-位点IRMA在抗原方面更经济，不太可能发生有害的变构反应，并且具有较低的零剂量反应（系统中总放射性的0.5-2%）。
A 2-site immunoradiometric assay (2-site IRMA) for human serum ferritin is carried out by reaction of the ferritin solution with a solid-phase anti(human ferritin), followed by a second reaction in which the insoluble product is incubated with purified, radioactively labeled anti(human ferritin). Unreacted labeled antibodies remain in solution and are washed away. As the amount of ferritin increases, the radioactivity in the solid-phase increases. Factors affecting the assay were evaluated including (a) concentration and stability of solid-phase antibody, (b) variation in temperature, reaction times, and reagent concentrations, (c) stability and storage of antibody coated tubes, (d) effect of tube washing cycles, (e) effect of serum proteins and anticoagulants, (f) organ specificity of ferritin. A paradoxical fall in dose-response was seen at high dose. Statistical analysis, dose interpolation, and automatic data processing were carried out by a generalization of the logit/log method and computer programs used in conventional radioimmunoassay. The dependence of the variance on the position on the dose-response curve is different than that seen in radioimmunoassay systems and is reflected in a greater effective assay range. 2-Site IRMA may also have advantages in reagent stability, specificity, antigen protection, and suitability for automation. The properties of 2-site IRMA are closely related to the usual IRMA assay system, but 2-site IRMA is more economical in antigen, is unlikely to be subject to deleterious allosteric reactions, and has a lower zero dose-response (0.5–2% of the total radioactivity in the system).