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首页> 外文期刊>BioFactors >Endogenous morphine-6-glucuronide (M6G) is present in the plasma of patients: Validation of a specific anti-M6G antibody for clinical and basic research
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Endogenous morphine-6-glucuronide (M6G) is present in the plasma of patients: Validation of a specific anti-M6G antibody for clinical and basic research

机译：患者血浆中存在内源性吗啡-6-葡糖醛酸（M6G）：针对临床和基础研究的特定抗M6G抗体的验证

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Endogenous morphine and its derivatives (morphine-6-glucuronide [M6G]; morphine-3-glucuronide [M3G]) are formed by mammalian cells from dopamine. Changes in the concentrations of endogenous morphine have been demonstrated in several pathologies (sepsis, Parkinson’s disease, etc.), and they might be relevant as pathological markers. While endogenous morphine levels are detectable using enzyme-linked immunosorbant assay (ELISA), mass spectrometry (MS) analysis was, so far, the only approach to detect and quantify M6G. This study describes the preparation of a specific anti-M6G rabbit polyclonal antibody and its validation. The specificity of this antibody was assessed against 30 morphine-related compounds. Then, a M6G-specific ELISA-assay was tested to quantify M6G in the plasma of healthy donors, morphinetreated, and critically ill patients. The antibody raised against M6G displays a strong affinity for M6G, codeine-6-glucuronide, and morphine-3-6-glucuronide, whereas only weak cross-reactivities were observed for the other compounds. Both M6G-ELISA and LC-MS/MS approaches revealed the absence of M6G in the plasma of healthy donors (controls, n=8). In all positive donors treated with morphine-patch (n=5), M6G was detected using both M6G-ELISA and LC-MS/MS analysis. Finally, in a study on critically ill patients with circulating endogenous morphine (n-26), LC-MS/MS analysis revealed that 73% of the positive-patients (19 of 26), corresponding to high M6G-levels in M6G-ELISA, contained M6G. In conclusion, we show that endogenous M6G can be found at higher levels than morphine in the blood of morphine-naive patients. With respect to the interest of measuring endogenous M6G in pathologies, we provide evidences that our ELISA procedure represents a powerful tool as it can easily and specifically detect endogenous M6G levels.
机译：内源性吗啡及其衍生物（吗啡-6-葡糖醛酸化物[M6G];吗啡-3-葡糖醛酸化物[M3G]）是由多巴胺的哺乳动物细胞形成的。内源性吗啡浓度的变化已在多种病理学（败血症，帕金森氏病等）中得到证实，它们可能与病理学指标有关。虽然可以使用酶联免疫吸附测定（ELISA）来检测内源性吗啡水平，但到目前为止，质谱（MS）分析是检测和定量M6G的唯一方法。这项研究描述了特异性抗M6G兔多克隆抗体的制备及其验证。针对30种吗啡相关化合物评估了该抗体的特异性。然后，测试了M6G特异性ELISA测定法，以量化健康供体，吗啡治疗和重症患者血浆中的M6G。产生的针对M6G的抗体对M6G，可待因-6-葡萄糖醛酸和吗啡-3-6-葡萄糖醛酸具有很强的亲和力，而其他化合物的交叉反应性很弱。 M6G-ELISA和LC-MS / MS方法均显示健康供体血浆中不存在M6G（对照组，n = 8）。在所有使用吗啡贴剂治疗的阳性供体中（n = 5），使用M6G-ELISA和LC-MS / MS分析检测到M6G。最后，在一项针对危重患者的循环内源性吗啡的研究中（n-26），LC-MS / MS分析显示73％的阳性患者（26人中的19人）对应于M6G-ELISA中较高的M6G水平，包含M6G。总之，我们表明，在未使用吗啡的患者血液中，内源性M6G的水平高于吗啡。关于在病理中测量内源性M6G的兴趣，我们提供的证据表明，我们的ELISA程序代表了一种强大的工具，因为它可以轻松，特异性地检测内源性M6G的水平。

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