Low-cost analytical assays enable accessible detection of clinically and environmentally important analytes; however, common enzyme-based assays suffer from high production and storage costs. Catalytically active synthetic materials serve as replacements for natural enzymes, but development of cost-effective, highly efficient synthetic strategies remains a challenge. Here, we utilized a facile synthesis for copper bipyridine coordination polymers (CuBpyCPs) and investigated structure-function relationships to achieve optimal catalytic properties for a glucose assay.
View Article and Find Full Text PDFMetal-organic frameworks (MOFs) are a class of porous nanomaterials that have been extensively studied as enzyme immobilization substrates. During in situ immobilization, MOF nucleation is driven by biomolecules with low isoelectric points. Investigation of how biomolecules control MOF self-assembly mechanisms on the molecular level is key to designing nanomaterials with desired physical and chemical properties.
View Article and Find Full Text PDFSingle-molecule-enzymology (SME) methods have enabled the observation of heterogeneous catalytic activities within a single enzyme population. Heterogeneous activity is hypothesized to originate from conformational changes in the enzyme that result from changes in the local environment leading to catalytically active substates. Here, we use SME to investigate the mechanisms of heterogeneous activity exhibited by tissue nonspecific alkaline phosphatase (TNSALP), which reveals two subpopulations with different catalytic activities.
View Article and Find Full Text PDFHumoral immunity to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) during acute infection and convalescence has been widely studied since March 2020. In this review, the authors summarize literature on humoral responses to SARS-CoV-2 antigens with a focus on spike, nucleocapsid, and the receptor-binding domain as targets of antibody responses. They highlight serologic studies during acute SARS-CoV-2 infection and discuss the clinical relevance of antibody levels in COVID-19 progression.
View Article and Find Full Text PDFEnzymes can be used as biomarkers for a variety of diseases. However, profiling enzyme activity in clinical samples is challenging due to the heterogeneity in enzyme activity, and the low abundance of the target enzyme in biofluids. Single-molecule methods can overcome these challenges by providing information on the distribution of enzyme activities in a sample.
View Article and Find Full Text PDFBACKGROUNDWeeks after SARS-CoV-2 infection or exposure, some children develop a severe, life-threatening illness called multisystem inflammatory syndrome in children (MIS-C). Gastrointestinal (GI) symptoms are common in patients with MIS-C, and a severe hyperinflammatory response ensues with potential for cardiac complications. The cause of MIS-C has not been identified to date.
View Article and Find Full Text PDFSevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2) proteins were measured in longitudinal plasma samples collected from 13 participants who received two doses of mRNA-1273 vaccine. Eleven of 13 participants showed detectable levels of SARS-CoV-2 protein as early as day 1 after first vaccine injection. Clearance of detectable SARS-CoV-2 protein correlated with production of immunoglobulin G (IgG) and immunoglobulin A (IgA).
View Article and Find Full Text PDFSensitive assays are essential for the accurate identification of individuals infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Here, we report a multiplexed assay for the fluorescence-based detection of seroconversion in infected individuals from less than 1 µl of blood, and as early as the day of the first positive nucleic acid test after symptom onset. The assay uses dye-encoded antigen-coated beads to quantify the levels of immunoglobulin G (IgG), IgM and IgA antibodies against four SARS-CoV-2 antigens.
View Article and Find Full Text PDFMeasurements of multiple biomolecules within the same biological sample are important for many clinical applications to enable accurate disease diagnosis or classification. These disease-related biomarkers often exist at very low levels in biological fluids, necessitating ultrasensitive measurement methods. Single-molecule arrays (Simoa), a bead-based digital enzyme-linked immunosorbent assay, is the current state of the art for ultrasensitive protein detection and can detect sub-femtomolar protein concentrations, but its ability to achieve high-order multiplexing without cross-reactivity remains a challenge.
View Article and Find Full Text PDFJ Am Chem Soc
September 2020
Enzyme assays are important for many applications including clinical diagnostics, functional proteomics, and drug discovery. Current methods for enzymatic activity measurement often suffer from low analytical sensitivity. We developed an ultrasensitive method for the detection of enzymatic activity using Single Molecule Arrays (eSimoa).
View Article and Find Full Text PDFThe COVID-19 pandemic continues to infect millions of people worldwide. In order to curb its spread and reduce morbidity and mortality, it is essential to develop sensitive and quantitative methods that identify infected individuals and enable accurate population-wide screening of both past and present infection. Here we show that Single Molecule Array assays detect seroconversion in COVID-19 patients as soon as one day after symptom onset using less than a microliter of blood.
View Article and Find Full Text PDFDJ-1, a 20.7 kDa protein, is overexpressed in people who have bladder cancer (BC). Its elevated concentration in urine allows it to serve as a marker for BC.
View Article and Find Full Text PDFProtein-metal-organic frameworks (p-MOFs) are a prototypical example of how synthetic biological hybrid systems can be used to develop next-generation materials. Controlling p-MOF formation enables the design of hybrid materials with enhanced biological activity and high stability. However, such control is yet to be fully realized due to an insufficient understanding of the governing nucleation and growth mechanisms in p-MOF systems.
View Article and Find Full Text PDFACS Appl Mater Interfaces
October 2018
A wet chemical process involving two electrodeposition steps followed by a solution casting step, the "EESC" process, is described for the fabrication of electroluminescent, radial junction wires. EESC is demonstrated by assembling three well-studied nanocrystalline (or amorphous) materials: Au, CdSe, and poly(3,4-ethylenedioxythiophene):polystyrene sulfonate (PEDOT:PSS). The tri-layered device architecture produced by EESC minimizes the influence of an electrically resistive CdSe emitter layer by using a highly conductive gold nanowire that serves as both a current collector and a negative electrode.
View Article and Find Full Text PDFA new type of chemiresistor, the impedance-transduced chemiresistor (ITCR), is described for the rapid analysis of glucose. The ITCR exploits porous, high surface area, fluorine-doped carbon nanofibers prepared by electrospinning of fluorinated polymer nanofibers followed by pyrolysis. These nanofibers are functionalized with a boronic acid receptor and stabilized by Nafion to form the ITCR channel for glucose detection.
View Article and Find Full Text PDFThe virus bioresistor (VBR) is a chemiresistor that directly transfers information from virus particles to an electrical circuit. Specifically, the VBR enables the label-free detection of a target protein that is recognized and bound by filamentous M13 virus particles, each with dimensions of 6 nm ( w) × 1 μm ( l), entrained in an ultrathin (∼250 nm) composite virus-polymer resistor. Signal produced by the specific binding of virus to target molecules is monitored using the electrical impedance of the VBR: The VBR presents a complex impedance that is modeled by an equivalent circuit containing just three circuit elements: a solution resistance ( R), a channel resistance ( R), and an interfacial capacitance ( C).
View Article and Find Full Text PDFACS Appl Mater Interfaces
November 2017
Pd based alloy materials with hollow nanostructures are ideal hydrogen (H) sensor building blocks because of their double-H sensing active sites (interior and exterior side of hollow Pd alloy) and fast response. In this work, for the first time, we report a simple fabrication process for preparing hollow Pd-Ag alloy nanowires (Pd@Ag HNWs) by using the electrodeposition of lithographically patterned silver nanowires (NWs), followed by galvanic replacement reaction (GRR) to form palladium. By controlling the GRR time of aligned Ag NWs within an aqueous Pd-containing solution, the compositional transition and morphological evolution from Ag NWs to Pd@Ag HNWs simultaneously occurred, and the relative atomic ratio between Pd and Ag was controlled.
View Article and Find Full Text PDFThe oxygen, O, in air interferes with the detection of H by palladium (Pd)-based H sensors, including Pd nanowires (NWs), depressing the sensitivity and retarding the response/recovery speed in air-relative to N or Ar. Here, we describe the preparation of H sensors in which a nanofiltration layer consisting of a Zn metal-organic framework (MOF) is assembled onto Pd NWs. Polyhedron particles of Zn-based zeolite imidazole framework (ZIF-8) were synthesized on lithographically patterned Pd NWs, leading to the creation of ZIF-8/Pd NW bilayered H sensors.
View Article and Find Full Text PDFThe influence of hexamethylenetetraamine (HMTA) on the morphology of δ-MnO and its properties for electrical energy storage are investigated-specifically for ultrathick δ-MnO layers in the micron scale. Planar arrays of gold@δ-MnO, core@shell nanowires, were prepared by electrodeposition with and without the HMTA and their electrochemical properties were evaluated. HMTA alters the MnO in three ways: First, it creates a more open morphology for the MnO coating, characterized by "petals" with a thickness of 6 to 9 nm, rather than much thinner δ-MnO sheets seen in the absence of HMTA.
View Article and Find Full Text PDFThe label-free detection of human serum albumin (HSA) in aqueous buffer is demonstrated using a simple, monolithic, two-electrode electrochemical biosensor. In this device, both millimeter-scale electrodes are coated with a thin layer of a composite containing M13 virus particles and the electronically conductive polymer poly(3,4-ethylenedioxy thiophene) or PEDOT. These virus particles, engineered to selectively bind HSA, serve as receptors in this biosensor.
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