Abstract
BackgroundAnti-myeloperoxidase antibody (anti-MPO) is an important biomarker for anti-neutrophil cytoplasm antibody (ANCA)-associated vasculitides (AAVs). However, the complicated operation procedures and insufficient sensitivity of conventional anti-MPO detection methods limit their application in monitoring efficacy of AAVs in clinical diagnosis. Herein, a dual amplified electrochemiluminescence (ECL) immunosensor based on multi-function PtCo nanozymes/CdS nanocrystals@graphene oxide (PtCo/CdS@GO) luminophores and K2S2O8/H2O2 coreactants has been fabricated for ultrasensitive detection of anti-MPO.ResultsPtCo/CdS@GO luminophores as novel signal amplification labels and nanocarriers to load rabbit anti-mouse IgG were synthesized by co-doping with Pt and Co nanozymes simultaneously with several considerable advantages, including astonishing peroxidase-like catalytic activity, high-efficiency luminescence performance and superior stability in aqueous solutions. Meanwhile, upon the K2S2O8/H2O2 coreactants system, benefiting from the efficient peroxidase-like activity of the PtCo/CdS@GO toward H2O2, massive of transient reactive intermediates could react with K2S2O8, thus obtaining higher ECL emission. Therefore, the developed ECL immunosensor for anti-MPO detection displayed good analytical performance with good concentration linearity in the range of 0.02 to 1000 pg/mL and low detection limit down to 7.39 fg/mL.ConclusionsThe introduction of multi-function PtCo/CdS@GO luminophores into the established ECL immunoassay not only was successfully applied for specific detection of anti-MPO in clinical serum samples, but also provided a completely new concept to design other high-performance luminophores. Meaningfully, the ECL immunoassay strategy held wide potential for biomarkers detection in clinical diagnosis.Graphic abstract
Highlights
Anti-myeloperoxidase antibody is an important biomarker for anti-neutrophil cytoplasm antibody (ANCA)-associated vasculitides (AAVs)
EDS elemental analysis was used to Characterization of Au nanoparticles functionalized MoS2 (Au@MoS2) nanosheets MoS2 nanosheets is a kind of layered nanomaterials that can stabilize metal NPs and expand their functions due to its high surface area and two-dimensional ultrathin atomic layer structure [31]
When horseradish peroxidase (HRP) was added to TMB/H2O2 system, a highest absorbance slope was obtained, which was significantly higher than that of Fe3O4 and slightly higher than that of PtCo/CdS@GO luminophores (CdS)@GO luminophores (PtCo)/CdS@graphene oxide (GO) and PtCo/rGO. These results showed that the PtCo/CdS@GO possessed a comparable catalytic activity with HRP
Summary
Anti-myeloperoxidase antibody (anti-MPO) is an important biomarker for anti-neutrophil cytoplasm antibody (ANCA)-associated vasculitides (AAVs). The complicated operation procedures and insufficient sensitivity of conventional anti-MPO detection methods limit their application in monitoring efficacy of AAVs in clinical diagnosis. According to the International Consensus on ANCA Testing in Eosinophilic Granulomatosis with Polyangiitis and the Revised 2017 international consensus on testing of ANCAs in granulomatosis with polyangiitis and microscopic polyangiitis, indirect immunofluorescence (IIF) and enzyme-linked immunosorbent assays (ELISA) have been used to initially screen and verify anti-MPO [3, 4]. These conventional anti-MPO detection methods usually suffer from complicated operation procedures, insufficient sensitivity and specificity [5, 6].
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