Criteria of evaluation and of interpretation of Sepharose drug IgE‐RIA to anaesthetic drugs
Criteria of evaluation and of interpretation of Sepharose drug IgE‐RIA to anaesthetic drugs
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Sepharose药物IgE-RIA对麻醉药物的评价和解释标准
DOI：
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发表时间：
1999
期刊：
影响因子：
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通讯作者：
M. Laxenaire
中科院分区：
文献类型：
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作者：
J. Guéant;E. Mata;F. Namour;A. Romano;I. Aimone;Gisèle Kanny;D. A. Moneiet;M. Laxenaire
血清中抗药物特异性IgE的检测通常是通过夹心式免疫分析进行的，即首先将血清IgE吸附到反应相，然后通过结合抗IgE示踪剂进行定量。制备新的药物反应相需要仔细建立不同的验证步骤：1)对照血清的阳性标准2)与可溶性药物的竞争抑制分析，其中应包括抑制常数的测定，而不是单个抑制百分比的估计，特别是当分析是为了识别决定因素时3)估计IgE与固相的非特异性结合，包括疏水结合。竞争抑制作用取决于竞争试剂和试管中IgE的浓度，以及固相结合活性药物的浓度。我们通过Dixon检验来计算竞争对手的抑制常数(Ki)，从而改进了抑制试验。对12例对肌松药过敏反应的患者进行了6种不同肌松药的KI测定。数值范围在1.5纳米到2.5微米之间。这证实了患者之间药物IgE交叉反应的高度异质性。在12名患者中，有8名患者涉案药物的Ki值最低(亲和力最高)。与经典的抑制法相比，该方法与临床数据的相关性更好。接近四元离子的疏水环境似乎是必要的，以允许IgE与肌肉松弛药结合。相反，在Tiemonium中，一个距离四元离子约3A的羟基可以解释为什么这种分子具有高KI(微米)。总之，在分子识别研究中，应该建议测定可溶性药物和相关化合物的抑制常数，以补充仅基于半抗原抑制试验的实验。
The detection of antidrug specific IgE in serum is usually performed by a sandwich-type immunoassay in which the serum IgE is first adsorbed to a reactive phase and subsequently quantified via the binding of an anti-IgE tracer. The preparation of a new drug-reactive phase requires one to establish carefully different steps of validation: 1) criteria of positivity of control sera 2) competitive inhibition assays with the soluble drug, which should include the determination of the inhibition constant rather than estimation of a single inhibition percentage, especially when the assay is performed for the identification of determinants 3) estimation of nonspecific binding of IgE to the solid phase, including hydrophobic binding. The competitive inhibition depends on the concentration of the competitor and of IgE in the test-tube and the concentration of reactive drug bound to the solid phase. We have improved the inhibition assay by performing the Dixon test for calculating the inhibition constant (Ki) of the competitor. The Ki of six different muscle relaxants was determined in 12 patients who experienced an anaphylactic reaction to muscle relaxants. The values ranged between 1.5 nM and 2.5 microM. This confirmed the great heterogeneity of drug IgE cross-reactivity among patients. The Ki value of the incriminated drug was the lowest (affinity, the highest) in eight of the 12 patients. It was better correlated to clinical data than the classical inhibition assay. A hydrophobic environment seemed to be necessary, close to the quaternary ion, to allow IgE binding to the muscle relaxant. By contrast, in tiemonium, a hydroxyl group present at a distance of about 3 A from the quaternary ion may explain why this molecule had a high Ki (microM). In conclusion, it should be recommended, in molecular-recognition studies, that the inhibition constant of the soluble drug and of the related compounds be determined to complement the experiments based only on hapten inhibition assays.