2D Graphene Oxide/Fe-MOF Nanozyme Nest with Superior Peroxidase-Like Activity and Its Application for Detection of Woodsmoke Exposure Biomarker.

Emerging nanomaterials like nanozymes have recently been applied for the immunoassay-based detection of biomarkers. However, the inferior catalytic activity and low water solubility of nanozymes remain as the major limitations compared to natural enzymes. To overcome these limitations, we successfully synthesized a superior nanozyme with a structure of enriched 2D catalytic interface, namely Nanozyme Nest, which was composed of Fe-based metal-organic frameworks (Fe-MOF) and graphene oxide (GO). Then, we applied it in an ultrasensitive enzyme-linked immunosorbent assay (ELISA) for the detection of benzo[a]pyrene-7,8-diol-9,10-epoxide-DNA adduct (BPDE-DNA), which is a metabolite of benzo[a]pyrene (BP) and used as a typical biomarker of woodsmoke exposure in human blood. The Nanozyme Nest features amplified peroxidase-like catalytic ability from graphene and Fe-MOF due to their large surface area and abundant active sites. By using the proposed Nanozyme Nest based ultrasensitive ELISA, the BPDE-DNA could be detected at a level as low as 0.268 ng/mL, and the obtained sensitivity was much higher than most of the widely used methods. Our work provides a novel strategy to design ultrasensitive immunosensors with advantages of amplified catalytic activity and improved wa-ter solubility compared to classic nanozymes. This illustrates the promising applications of the Nanozyme Nest based immunosensors in point-of-care settings to conveniently detect exposures and diagnose diseases.

[1]  A. Hartwig,et al.  BPDE-induced genotoxicity: relationship between DNA adducts, mutagenicity in the in vitro PIG-A assay, and the transcriptional response to DNA damage in TK6 cells , 2017, Archives of Toxicology.

[2]  Yuehe Lin,et al.  Recent Advances in Biosensors for Detecting Cancer-Derived Exosomes. , 2019, Trends in biotechnology.

[3]  Chengzhou Zhu,et al.  Electrochemical Sensors and Biosensors Based on Nanomaterials and Nanostructures , 2014, Analytical chemistry.

[4]  G. Molnár,et al.  Magnetite Fe3 O4 Has no Intrinsic Peroxidase Activity, and Is Probably not Involved in Alzheimer's Oxidative Stress. , 2018, Angewandte Chemie.

[5]  Ya Li Liu,et al.  A nanosized metal-organic framework of Fe-MIL-88NH₂ as a novel peroxidase mimic used for colorimetric detection of glucose. , 2013, The Analyst.

[6]  Wei Wen,et al.  Recent progress in biosensors based on organic-inorganic hybrid nanoflowers. , 2018, Biosensors & bioelectronics.

[7]  Junbo Chen,et al.  Binding-Induced Molecular Amplifier as a Universal Detection Platform for Biomolecules and Biomolecular Interaction. , 2018, Analytical chemistry.

[8]  Jixuan Liu,et al.  Smartphone-based analytical biosensors. , 2018, The Analyst.

[9]  Ying Wang,et al.  Discovering the enzyme mimetic activity of metal-organic framework (MOF) for label-free and colorimetric sensing of biomolecules. , 2016, Biosensors & bioelectronics.

[10]  Qin Xu,et al.  Nickel metal-organic framework 2D nanosheets with enhanced peroxidase nanozyme activity for colorimetric detection of H2O2. , 2018, Talanta.

[11]  Li Fa-sheng,et al.  Quantitative analysis of 3-OHB[a]P and (+)-anti-BPDE as biomarkers of B[a]P exposure in rats. , 2016, Biomedical chromatography : BMC.

[12]  I. Willner,et al.  Stimuli‐Responsive DNA‐Functionalized Metal–Organic Frameworks (MOFs) , 2017, Advanced materials.

[13]  E. Pelletier,et al.  Subpicogram determination of (+)-anti-benzo[a]pyrene diol-epoxide adducts in fish albumin and globin by high-performance liquid chromatography with fluorescence detection , 2001 .

[14]  G. Speit,et al.  Further characterization of benzo[a]pyrene diol-epoxide (BPDE)-induced comet assay effects. , 2016, Mutagenesis.

[15]  Chengzhou Zhu,et al.  Nanozyme Enhanced Colorimetric Immunoassay for Naked-Eye Detection of Salmonella Enteritidis , 2018, Journal of Analysis and Testing.

[16]  Kangbing Wu,et al.  Strategy for Highly Sensitive Electrochemical Sensing: In Situ Coupling of a Metal-Organic Framework with Ball-Mill-Exfoliated Graphene. , 2019, Analytical chemistry.

[17]  Mohammad Hasanzadeh,et al.  Early stage screening of breast cancer using electrochemical biomarker detection , 2017 .

[18]  Dan Du,et al.  Graphene-like 2D nanomaterial-based biointerfaces for biosensing applications. , 2017, Biosensors & bioelectronics.

[19]  Dan Du,et al.  Functionalized graphene oxide as a nanocarrier in a multienzyme labeling amplification strategy for ultrasensitive electrochemical immunoassay of phosphorylated p53 (S392). , 2011, Analytical chemistry.

[20]  Yuehe Lin,et al.  Single-Atom Nanozyme Based on Nanoengineered Fe-N-C Catalyst with Superior Peroxidase-Like Activity for Ultrasensitive Bioassays. , 2019, Small.

[21]  Junfa Yin,et al.  Fluorescently imaged particle counting immunoassay for sensitive detection of DNA modifications. , 2010, Analytical chemistry.

[22]  Dan Du,et al.  A 3D-Printed, Portable, Optical-Sensing Platform for Smartphones Capable of Detecting the Herbicide 2,4-Dichlorophenoxyacetic Acid. , 2017, Analytical chemistry.

[23]  Na Liu,et al.  Determination of 3-OHB[a]P and (+)-anti-BPDE in Rats Blood and Brain Tissue of B[a]P Exposure by HPLC with Fluorescence Detection , 2015, Chromatographia.

[24]  X. Le,et al.  Quantum dots enhanced ultrasensitive detection of DNA adducts. , 2009, Analytical chemistry.

[25]  Gérard Férey,et al.  Porous metal-organic-framework nanoscale carriers as a potential platform for drug delivery and imaging. , 2010, Nature materials.

[26]  C. Winterbourn.  Toxicity of iron and hydrogen peroxide: the Fenton reaction. , 1995, Toxicology letters.

[27]  Nian-Lu Li,et al.  A novel sandwiched electrochemiluminescence immunosensor for the detection of carcinoembryonic antigen based on carbon quantum dots and signal amplification. , 2017, Biosensors & bioelectronics.

[28]  I. Weinstein,et al.  Quantitation of benzo(a)pyrene-deoxyguanosine adducts by radioimmunoassay. , 1980, Cancer research.

[29]  C. Téllez,et al.  Fabrication of ultrathin films containing the metal organic framework Fe-MIL-88B-NH2 by the Langmuir-Blodgett technique , 2015 .

[30]  David E. Williams,et al.  Point of care diagnostics: status and future. , 2012, Analytical chemistry.

[31]  Weihong Tan,et al.  Ultrasensitive DNA detection using highly fluorescent bioconjugated nanoparticles. , 2003, Journal of the American Chemical Society.

[32]  Xiaogang Qu,et al.  Carbon Nanozymes: Enzymatic Properties, Catalytic Mechanism, and Applications. , 2018, Angewandte Chemie.

[33]  X. Le,et al.  Binding-Induced DNA Dissociation Assay for Small Molecules: Sensing Aflatoxin B1. , 2018, ACS sensors.

[34]  Joseph Wang.  Nanomaterial-based amplified transduction of biomolecular interactions. , 2005, Small.

[35]  W. S. Hummers,et al.  Preparation of Graphitic Oxide , 1958 .

[36]  M. Weinfeld,et al.  Detection of benzo(a)pyrene diol epoxide‐DNA adducts in mononuclear white blood cells by CE immunoassay and its application to studying the effect of glutathione depletion , 2009, Electrophoresis.

[37]  T. E. Reed,et al.  Detection of benzo(a)pyrene diol epoxide-DNA adducts in sperm of men exposed to cigarette smoke. , 1999, Fertility and sterility.

[38]  Junxing Hao,et al.  Potential-Tunable Metal–Organic Frameworks: Electrosynthesis, Properties, and Applications for Sensing of Organic Molecules , 2019, The Journal of Physical Chemistry C.

[39]  C. Seymour,et al.  Point-of-care sensors for the management of sepsis , 2018, Nature Biomedical Engineering.

[40]  Dan Du,et al.  Smart phone based immunosensor coupled with nanoflower signal amplification for rapid detection of Salmonella Enteritidis in milk, cheese and water , 2017 .

[41]  Yuehe Lin,et al.  Unprecedented peroxidase-mimicking activity of single-atom nanozyme with atomically dispersed Fe-Nx moieties hosted by MOF derived porous carbon. , 2019, Biosensors & bioelectronics.

[42]  P. Tchounwou,et al.  A Review on Graphene-Based Nanomaterials in Biomedical Applications and Risks in Environment and Health , 2018, Nano-micro letters.

[43]  Dawei Feng,et al.  Zirconium-metalloporphyrin PCN-222: mesoporous metal-organic frameworks with ultrahigh stability as biomimetic catalysts. , 2012, Angewandte Chemie.

[44]  Dan Du,et al.  A portable smart-phone device for rapid and sensitive detection of E. coli O157:H7 in Yoghurt and Egg. , 2018, Biosensors & bioelectronics.

[45]  Ki‐Hyun Kim,et al.  TCNQ-doped Cu-metal organic framework as a novel conductometric immunosensing platform for the quantification of prostate cancer antigen , 2017 .

[46]  S. Kyrtopoulos,et al.  Detection and quantitation of benzo[a]pyrene-derived DNA adducts in mouse liver by liquid chromatography-tandem mass spectrometry: comparison with 32P-postlabeling. , 2006, Chemical research in toxicology.

[47]  Q. Wei,et al.  3D Nanostructured Palladium-Functionalized Graphene-Aerogel-Supported Fe3O4 for Enhanced Ru(bpy)32+-Based Electrochemiluminescent Immunosensing of Prostate Specific Antigen. , 2017, ACS applied materials & interfaces.

[48]  A. Tang,et al.  Standardized assays for determining the catalytic activity and kinetics of peroxidase-like nanozymes , 2018, Nature Protocols.

[49]  Xiaogang Qu,et al.  Nanozymes: Classification, Catalytic Mechanisms, Activity Regulation, and Applications. , 2019, Chemical reviews.

[50]  William W. Chen,et al.  Classic and contemporary approaches to modeling biochemical reactions. , 2010, Genes & development.

[51]  Shun Mao,et al.  Metal–Organic Framework-Based Sensors for Environmental Contaminant Sensing , 2018, Nano-Micro Letters.

[52]  V. Boddi,et al.  Benzo(a)pyrene diolepoxide (BPDE)-DNA adduct levels in leukocytes of smokers in relation to polymorphism of CYP1A1, GSTM1, GSTP1, GSTT1, and mEH. , 2004, Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology.

[53]  Junhua Song,et al.  Metal–organic frameworks-based catalysts for electrochemical oxygen evolution , 2019, Materials Horizons.

[54]  X. Le,et al.  Antibody-Bridged Beacon for Homogeneous Detection of Small Molecules. , 2018, Analytical chemistry.

[55]  Wenjun Yang,et al.  Nanoencapsulated microcrystalline particles for superamplified biochemical assays. , 2002, Analytical chemistry.

[56]  Kangbing Wu,et al.  Tunable Electrochemistry of Electrosynthesized Copper Metal–Organic Frameworks , 2018 .

[57]  V. Chen,et al.  Metal-Organic-Framework-Based Enzymatic Microfluidic Biosensor via Surface Patterning and Biomineralization. , 2018, ACS applied materials & interfaces.