New high mobility group box 1 assay system
New high mobility group box 1 assay system
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DOI：
10.1016/j.cca.2006.04.016
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发表时间：
2006-10-01
影响因子：
5
通讯作者：
Maruyama, Ikuro
中科院分区：
文献类型：
--
作者：
Yamada, Shingo;Yakabe, Keiko;Maruyama, Ikuro
背景：使用化学发光底物已开发出高灵敏度的夹心ELISA方法。高迁移率族蛋白1(HMGB1)在多种炎症性疾病中起重要作用，并可能参与动脉粥样硬化的形成。方法：制备抗人MGB1多肽序列(KPDAAKKGVV-KAEK)的抗人MGB1单抗和抗肽多克隆抗体(KPDAAKKGVV-KAEK)，并以其为显色底物(TMBZ)和化学发光底物(PS-ATTO)构建夹心ELISA方法。以高纯度人HMGB1为标准品，测定高敏C反应蛋白并与HMGB1进行比较。结果：建立的方法具有良好的检测性能，批内、批间变异系数均为10%，化学发光法的检出限为0.3mg/L，显色底物法的检出限为1mg/L。检测急性冠脉综合征(ACS)患者血清HMGB1水平。以入院时HMGB1为临界值时，HMGB I异常的急性冠脉综合征患者出院后1个月内发生急性心脏事件的风险显著高于HMGB I正常者(P<；0.0001)。提示HMGB1作为急性预后标记物的有效性。结论：该方法操作简便，适合在医院实验室和大规模人群筛查中使用。急性冠脉综合征患者血清中可检测到HMGB1，血清HMGBI浓度可作为判断患者预后的指标。(C)2006爱思唯尔B.V.保留所有权利。
Background: High-sensitivity sandwich ELISA methods have been developed using chemiluminescent substrates. HMGB1 (high mobility group box 1) protein has been shown to play a critical role in several inflammatory diseases and it may be involved in the development of atherosclerosis.Methods: Anti-human MGB1 monoclonal antibodies and anti-peptide polyclonal antibodies against the peptide sequence (KPDAAKKGVV-KAEK) with high antigenicity and different from the sequence of HMGB2 were developed, and the antibodies were used to construct sandwich ELISA methods with a chromogenic substrate (TMBZ) and a chemiluminescent substrate (PS-atto). Highly purified human HMGB1 was used as a standard material and high-sensitivity CRP was measured to compare with HMGB1.Results: The analytical characteristics of the ELISA method we developed were validated inter-assay and intra-assay CVs were < 10%, and the detection limit was 0.3 mu g/l by the chemiluminescent method and 1 mu g/l with the chromogenic substrates. HMGB1 was detected in the serum of patients with acute coronary syndrome (ACS). When a cut-off of 0.6 mu g/l HMGB1 upon admission to the intensive care unit (ICU) was used, the risk of developing an acute cardiac event within I month after discharge of ACS patients with an abnormal HMGB I was significantly higher than for the patients with normal values (P < 0.0001). The usefulness of HMGB1 as an acute prognostic marker was suggested.Conclusions: The assay is easy to perform and suitable for use in the hospital laboratory and for screening large populations. HMGB1 is detectable in the serum of ACS patients and that the serum concentration of HMGBI may be a prognostic indicator in ACS patients. (c) 2006 Elsevier B.V. All rights reserved.