Erhard M H, Kühlmann R, Szinicz L, Lösch U
Institut für Physiologie, Physiologische Chemie und Ernährungsphysiologie, Tierärztliche Fakultät, Universität München, Federal Republic of Germany.
Arch Toxicol. 1990;64(7):580-5. doi: 10.1007/BF01971838.
The development of a specific and sensitive immunologic ELISA detection system for methylphosphonoflouridic acid. 1,2,2-trimethylpropylester (soman) by the use of monoclonal antibodies (MAbs) is described. The monoclonal antibodies F71D7, F71H10, F71B12 and F71H9 originally produced against the soman derivative methyl phosphonic acid, p-aminophenyl 1,2,2-trimethylpropyldiester (MATP) also reacted with soman in a previously developed, direct competitive ELISA. After optimizing the ELISA system by varying the reaction mixture and the solvents for the organophosphate, 5.0 x 10(-7) mol/l soman (80% purity), e.g. 2.5 ng or 2 ng pure soman per 25 microliters test buffer, could be detected after a total test duration of 40 min. A shortening of the incubation time to 10 min resulted in a drop of sensitivity to 1.8 x 10(-6) mol/l soman. Various alcohols which may be used as extraction media for soman from various materials (isopropanol, ethanol and methanol) were shown to inhibit peroxidase activity and thereby reduce the sensitivity of the test. However, the influence of alcohols decreased with the shortening of incubation time. All monoclonal antibodies showed little cross reactivity to sarin and no cross reactivity to tabun and VX. Judging on the reactivity of the MAbs with MATP and soman oxidazed by 1,2-dihydrobenzol, some reactivity with some other (non-toxic) soman analogues containing the same pinacolyl group can be expected. There was no evidence for stereoselectivity of the MAbs tested. Finally, soman could be detected in different biological samples like human serum, goat serum, rabbit serum, chicken serum, milk, and tap water in concentrations between 1.3 x 10(-6) and 2.0 x 10(-6) mol/l.
描述了一种通过使用单克隆抗体(MAb)开发的用于甲基膦酰氟代异丁酸1,2,2-三甲基丙酯(梭曼)的特异性和灵敏性免疫酶联免疫吸附测定(ELISA)检测系统。最初针对梭曼衍生物甲基膦酸对氨基苯基1,2,2-三甲基丙酯(MATP)产生的单克隆抗体F71D7、F71H10、F71B12和F71H9,在先前开发的直接竞争ELISA中也与梭曼发生反应。通过改变反应混合物和有机磷酸酯的溶剂对ELISA系统进行优化后,在总测试时长40分钟后,可检测到5.0×10⁻⁷摩尔/升的梭曼(纯度80%),例如每25微升测试缓冲液中含有2.5纳克或2纳克纯梭曼。将孵育时间缩短至10分钟会导致灵敏度降至1.8×10⁻⁶摩尔/升的梭曼。各种可用作从各种材料中提取梭曼的萃取介质的醇类(异丙醇、乙醇和甲醇)显示会抑制过氧化物酶活性,从而降低测试的灵敏度。然而,随着孵育时间的缩短,醇类的影响会降低。所有单克隆抗体对沙林几乎没有交叉反应性,对塔崩和VX也没有交叉反应性。根据单克隆抗体与MATP和被1,2 - 二氢苯氧化的梭曼的反应性判断,可以预期与一些其他含有相同频哪醇基的(无毒)梭曼类似物有一些反应性。所测试的单克隆抗体没有立体选择性的证据。最后,在人血清、山羊血清、兔血清、鸡血清、牛奶和自来水中能够检测到浓度在1.3×10⁻⁶至2.0×10⁻⁶摩尔/升之间的梭曼。