Graphene-Oxide-Loaded Superparamagnetic Iron Oxide Nanoparticles for Ultrasensitive Electrocatalytic Detection of MicroRNA

We report the electrocatalytic activity of a new class of superparamagnetic nanoparticles, graphene-oxide-loaded iron oxide (GO/IO hybrid material), towards the reduction of ruthenium hexaammine(III) chloride (Ru(NH3)6]3+, RuHex). Leveraging the electrocatalytic activity of the GO/IO hybrid material and the signal enhancement capacity of [Ru(NH3)6]3+/[Fe(CN)6]3 in an electrocatalytic cycle, an ultrasensitive and specific electrochemical sensor was developed for the detection of cancerrelated microRNA (miRNA). Using the direct affinity interaction between RNA and graphene oxide, magnetically isolated and purified target miRNA were directly adsorbed onto a screenprinted electrode modified with the GO/IO hybrid material. The detection was enabled by chronocoulometric (CC) readout of charge-compensating [Ru(NH3)6]3+ followed by an enhancement in CC charge display through the Ru(NH3)6]3+ /[Fe(CN)6]3 system. We demonstrate an excellent limit of detection of 1.0 fM by accurately detecting miR-21 in synthetic samples and showcase its clinical utility in ovarian cancer cell lines with high sensitivity (ten cells) and good reproducibility (%RSD= \u3c5%, for n=3)

Autoantibodies as diagnostic and prognostic cancer biomarker: detection techniques and approaches

Autoantibodies produced by the patients’ own immune systems in response to foreign substances are emerging as an attractive biomarker for early detection of cancer. These serum immunobiomarkers are produced in large quantities despite the presence of very less amount of the corresponding antigens, and thus presenting themselves as a novel class of stable and minimally invasive disease biomarkers especially for cancer diagnosis. Although a plethora of research, including conventional molecular biology-based as well as cutting-edge optical and electrochemical strategies (biosensor), have been conducted to detect autoantibodies, most of these strategies are yet to be readily applicable in the off-laboratory settings at clinics. Herein, we detail the biogenesis, diagnostic, prognostic and therapeutic potential of autoantibodies as cancer biomarkers. With the particular emphasis on cutting-edge advances in electrochemistry, optical (surface plasmon resonance)and microfluidics techniques, this review entrusts the unmet needs and challenges of autoantibody detection approaches and provides a future perspective of the presented strategies. We believe this review can potentially guide the researchers towards the development of robust, reliable and sensitive detection strategies for tumor-associated autoantibodies and translation of these biomarkers to real clinical settings for diagnosis and prognosis of cancer

Nanoarchitecture frameworks for electrochemical miRNA detection

With revolutionary advances in next-generation sequencing, the human transcriptome has been comprehensively interrogated. These discoveries have highlighted the emerging functional and regulatory roles of a large fraction of RNAs suggesting the potential they might hold as stable and minimally invasive disease biomarkers. Although a plethora of molecular-biology- and biosensor-based RNA-detection strategies have been developed, clinical application of most of these is yet to be realized. Multifunctional nanomaterials coupled with sensitive and robust electrochemical readouts may prove useful in these applications. Here, we summarize the major contributions of engineered nanomaterials-based electrochemical biosensing strategies for the analysis of miRNAs. With special emphasis on nanostructure-based detection, this review also chronicles the needs and challenges of miRNA detection and provides a future perspective on the presented strategies

Synthesis, spectroscopic characterization, thermal and luminescent properties of new organosulfur-functionalized platinum(II) bis(alkenylarylalkynyl) complexes

A series of organosulfur-functionalized trans-platinum(II) bis(alkenylarylalkynyl) complexes, having one tolylthio moiety in each alkenyl backbone with general formula trans-[(PEt 3 ) 2 Pt{CC-Ar-CH=CH(SC 6 H 4 -CH 3 )} 2 ], (2a-2d), (where, Ar = phenylene, biphenylene, 2,5-dimethylphenylene, and 2,5-dimethoxyphenylene) were synthesized in good to excellent yields with good regioselectivity. As compared to the absorption band of trans-platinum(II) bis(alkynylarylalkynyl) complexes, we found that the position of the lowest energy absorption bands in the trans-platinum(II) bis(alkenylarylalkynyl) complexes were red-shifted, after the functionalization of the trans-platinum(II) bis(alkynylarylalkynyl) complexes with arylthiol. For all trans-platinum(II) complexes, the lowest energy absorption bands in the UV/Vis spectra, in chloroform solution, at room temperature, were observed in the range 362-394 nm, and under excitation at the wavelength of the absorption maximum exhibited the emission peak maximum at room temperature in the range 401-426 nm. The newly synthesized complexes are not exhibited phosphorescence at room temperature but are exhibited at low temperature, 77 K. All the new platinum(II) complexes have been fully characterized by spectroscopic analysis as well as elemental analysis, and the trans square-planar arrangement at the platinum centre has been confirmed by single-crystal X-ray diffraction study of complex 2a

Designed Patterning of Mesoporous Metal Films Based on Electrochemical Micelle Assembly Combined with Lithographical Techniques

Mesoporous noble metals and their patterning techniques for obtaining unique patterned structures are highly attractive for electrocatalysis, photocatalysis, and optoelectronics device applications owing to their expedient properties such as high level of exposed active locations, cascade electrocatalytic sites, and large surface area. However, patterning techniques for mesoporous substrates are still limited to metal oxide and silica films, although there is growing demand for developing techniques related to patterning mesoporous metals. In this study, the first demonstration of mesoporous metal films on patterned gold (Au) substrates, prefabricated using photolithographic techniques, is reported. First, different growth rates of mesoporous Au metal films on patterned Au substrates are demonstrated by varying deposition times and voltages. In addition, mesoporous Au films are also fabricated on various patterns of Au substrates including stripe and mesh lines. An alternative fabrication method using a photoresist insulating mask also yields growth of mesoporous Au within the patterning. Moreover, patterned mesoporous films of palladium (Pd) and palladium–copper alloy (PdCu) are demonstrated on the same types of substrates to show versatility of this method. Patterned mesoporous Au films (PMGF) shows higher electrochemically-active surface area (ECSA) and higher sensitivity toward glucose oxidation than nonpatterned mesoporous Au films (NMGF)

Quantification of gene-specific DNA methylation in oesophageal cancer via electrochemistry

Development of simple and inexpensive method for the analysis of gene-specific DNA methylation is important for the diagnosis and prognosis of patients with cancer. Herein, we report a relatively simple and inexpensive electrochemical method for the sensitive and selective detection of gene-specific DNA methylation in oesophageal cancer. The underlying principle of the method relies on the affinity interaction between DNA bases and unmodified gold electrode. Since the affinity trend of DNA bases towards the gold surface follows as adenine (A) > cytosine (C) > guanine (G)> thymine (T), a relatively larger amount of bisulfite-treated adenine-enriched unmethylated DNA adsorbs on the screen-printed gold electrodes (SPE-Au) in comparison to the guanine-enriched methylated sample. The methylation levels were (i.e., different level of surface attached DNA molecules due to the base dependent differential adsorption pattern) quantified by measuring saturated amount of charge-compensating [Ru(NH ) ] molecules in the surface-attached DNAs by chronocoulometry as redox charge of the [Ru(NH ) ] molecules quantitatively reflects the amount of the adsorbed DNA confined at the electrode surface. The assay could successfully distinguish methylated and unmethylated DNA sequences at single CpG resolution and as low as 10% differences in DNA methylation. In addition, the assay showed fairly good reproducibility (% RSD

Superparamagnetic nanoarchitectures for disease-specific biomarker detection

The detection of clinically relevant disease-specific biomolecules, including nucleic acids, circulating tumor cells, proteins, antibodies, and extracellular vesicles, has been indispensable to understand their functions in disease diagnosis and prognosis. Therefore, a biosensor for the robust, ultrasensitive, and selective detection of these low-abundant biomolecules in body fluids (blood, urine, and saliva) is emerging in current clinical research. In recent years, nanomaterials, especially superparamagnetic nanomaterials, have played essential roles in biosensing due to their intrinsic magnetic, electrochemical, and optical properties. However, engineered multicomponent magnetic nanoparticle-based current biosensors that offer the advantages of excellent stability in a complex biomatrix; easy and alterable biorecognition of ligands, antibodies, and receptor molecules; and unified point-of-care integration have yet to be achieved. This review introduces the recent advances in superparamagnetic nanostructures for electrochemical and optical biosensing for disease-specific biomarkers. This review emphasizes the synthesis, biofunctionalization, and intrinsic properties of nanomaterials essential for robust, ultrasensitive biosensing. With a particular emphasis on nanostructure-based electrochemical and optical detection of disease-specific biomarkers such as nucleic acids (DNA and RNA), proteins, autoantibodies, and cells, this review also chronicles the needs and challenges of nanoarchitecture-based detection. These summaries provide further insights for researchers to inspire their future work on the development of nanostructures for integrating into biosensing and devices for a broad field of applications in analytical sensing and in clinic

Gold-loaded nanoporous ferric oxide nanocubes for electrocatalytic detection of microRNA at attomolar level

A crucial issue in microRNA (miRNA) detection is the lack of sensitive method capable of detecting the low levels of miRNA in RNA samples. Herein, we present a sensitive and specific method for the electrocatalytic detection of miR-107 using gold-loaded nanoporous superparamagnetic iron oxide nanocubes (Au-NPFe 2 O 3 NC). The target miRNA was directly adsorbed onto the gold surfaces of Au-NPFe 2 O 3 NC via gold-RNA affinity interaction. The electrocatalytic activity of Au-NPFe 2 O 3 NC was then used for the reduction of ruthenium hexaammine(III) chloride (RuHex, [Ru(NH 3 ) 6 ] 3+ ) bound with target miRNA. The catalytic signal was further amplified by using the ferri/ferrocyanide [Fe(CN) 6 ] 3-/4- system. These multiple signal enhancement steps enable our assay to achieve the detection limit of 100 aM which is several orders of magnitudes better than most of the conventional miRNA sensors. The method was also successfully applied to detect miR-107 from cancer cell lines and a panel of tissue samples derived from patients with oesophageal squamous cell carcinoma with excellent reproducibility (% RSD = \u3c 5%, for n = 3) and high specificity. The analytical accuracy of the method was validated with a standard RT-qPCR method. We believe that our method has the high translational potential for screening miRNAs in clinical samples

Porous nanozymes: The peroxidase-mimetic activity of mesoporous iron oxide for the colorimetric and electrochemical detection of global DNA methylation

Nanomaterials (nanozymes) with peroxidase-mimetic activity have been widely used in biosensing platforms as low-cost, relatively stable and prevailing alternatives to natural enzymes. Herein, we report on the synthesis and application of the peroxidase-mimetic activity of mesoporous iron oxide (MIO) for the detection of global DNA methylation in colorectal cancer cell lines. The target DNA was extracted and denatured to get ssDNA followed by direct adsorption onto the surface of a bare screen-printed gold electrode (SPGE). A 5-methylcytosine antibody (5mC) functionalized nanomaterial (MIO-5mC) was then used to recognise the methylcytosine groups present on the SPGE. The MIO-5mC conjugates catalyse the TMB solution in the presence of hydrogen peroxide to give the colorimetric (i.e., naked-eye observation) and electrochemical detection of DNA methylation. The assay could successfully detect as low as 10% difference in the global DNA methylation level in synthetic samples and cell lines with good reproducibility and specificity (%RSD = <5%, for n = 3). This strategy avoids the use of natural enzyme horseradish peroxidase (HRP), traditional PCR based amplification and bisulfite treatment steps that are generally used in many conventional DNA methylation assays. We envisage that our assay could be a low-cost platform with great potential for genome-wide DNA methylation analysis in point-of-care applications.Griffith Sciences, School of Environment and ScienceFull Tex