PtCo nanocubes/reduced graphene oxide hybrids and hybridization chain reaction-based dual amplified electrochemiluminescence immunosensing of antimyeloperoxidase
PtCo nanocubes/reduced graphene oxide hybrids and hybridization chain reaction-based dual amplified electrochemiluminescence immunosensing of antimyeloperoxidase
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PtCo纳米立方体/还原氧化石墨烯杂化物和基于杂交链式反应的抗髓过氧化物酶双放大电化学发光免疫传感
DOI：
10.1016/j.bios.2019.111548
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发表时间：
2019
影响因子：
12.6
通讯作者：
Zhao Min
中科院分区：
文献类型：
--
作者：
Yang Wei;Peng Qiling;Guo Zhen;Wu Haiping;Ding Shijia;Chen Yongjian;Zhao Min
抗髓过氧化物酶（anti-MPO）被认为是抗中性粒细胞胞浆抗体（ANCA）相关性血管炎（AAV）的最重要的循环自身抗体之一。因此，高灵敏度检测抗MPO以监测AAV在临床诊断中的功效至关重要。本文将还原氧化石墨烯负载的PtCo纳米立方体杂化物（PtCo@rGO）与杂交链反应（HCR）相结合，构建了一种用于检测MPO的高灵敏度电化学发光免疫传感器。将N-（氨基丁基）-N-（乙基异鲁米诺）（ABEI）与阿霉素（Dox）交联制备的多个ECL发光体（Dox-ABEI）嵌入HCR的双链DNA产物中，实现了ECL发光体的有效固定，获得了较强的ECL发射。由于PtCo@rGO对H2 O2的高效催化活性，产生了大量的超氧自由基（O2●-），进一步与ABEI反应产生电致发光。因此，所设计的用于抗MPO检测的ECL免疫测定法在50 fg/mL至1 ng/mL的浓度变化范围内表现出优异的灵敏度，检测限为15.68 fg/mL。重要的是，本工作提出了一种灵敏度高、特异性好的无酶电化学发光免疫分析方法，用于蛋白质检测的临床诊断。
Antimyeloperoxidase (anti-MPO) is regarded as one of the most important circulating autoantibodies for anti-neutrophil cytoplasm antibody (ANCA)-associated vasculitides (AAVs). Hence, it is crucial for highly sensitive detection of anti-MPO to monitor efficacy of AAVs in clinical diagnosis. Herein, a highly sensitive electrochemiluminescence (ECL) immunosensor for anti-MPO detection was constructed by combining reduced graphene oxide-supported PtCo nanocubes hybrids (PtCo@rGO) with hybridization chain reaction (HCR) as signal amplification. Multiple ECL luminophores (Dox-ABEI) prepared by cross-linking of N-(aminobutyl)-N-(ethylisoluminol) (ABEI) and doxorubicin (Dox) were intercalated into dsDNA products of HCR, achieving the effective immobilization of ECL luminophores to obtain strong ECL emission. Benefiting from the efficient catalytic activity of PtCo@rGO toward H2O2, the massive the superoxide radical (O2●-) were generated to further react with ABEI for ECL emission. Thus, the designed ECL immunoassay for anti-MPO detection exhibited excellent sensitivity of a concentration variation from 50 fg/mL to 1 ng/mL and a detection limit of 15.68 fg/mL. Importantly, this work proposed an enzyme-free ECL immunoassay with high sensitivity, excellent specificity for protein detection in clinical diagnosis.