DJ-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. However, no biosensor for the detection of DJ-1
has been demonstrated. Here, we describe a virus bioresistor (VBR) capable of detecting DJ-1 in urine at a concentration
of 10 pM in 1 min. The VBR consists of a pair of
millimeter-scale gold electrodes that measure the electrical impedance
of an ultrathin (≈ 150–200 nm), two-layer polymeric
channel. The top layer of this channel (90–105 nm in thickness)
consists of an electrodeposited virus-PEDOT (PEDOT is poly(3,4-ethylenedioxythiophene))
composite containing embedded M13 virus particles that are engineered
to recognize and bind to the target protein of interest, DJ-1. The
bottom layer consists of spin-coated PEDOT–PSS (poly(styrenesulfonate)).
Together, these two layers constitute a current divider. We demonstrate
here that reducing the thickness of the bottom PEDOT–PSS layer
increases its resistance and concentrates the resistance drop of the
channel in the top virus-PEDOT layer, thereby increasing the sensitivity
of the VBR and enabling the detection of DJ-1. Large
signal amplitudes coupled with the inherent simplicity of the VBR sensor design result in high signal-to-noise (S/N > 100) and excellent sensor-to-sensor
reproducibility characterized by coefficients of variation in the
range of 3–7% across the DJ-1 binding curve down to a concentration
of 30 pM, near the 10 pM limit of detection (LOD), encompassing four
orders of magnitude in concentration.
The COVID-19 pandemic has resulted in over two million deaths worldwide. Despite efforts to fight the virus, the disease continues to overwhelm hospitals with severely ill patients.
Characterization of antibody response to SARS-CoV-2 is urgently needed to predict COVID-19 disease trajectories. Ineffective antibodies or antibody-dependent enhancement (ADE) could derail patient immune responses, for example. ELISA and coronavirus antigen microarray (COVAM) analysis epitope-mapped plasma from 86 COVID-19 patients. The experiments identified antibodies to a 21-residue epitope from nucleocapsid (termed Ep9) associated with severe disease, including ICU stay, requirement for ventilators, and death. Furthermore, anti-Ep9 antibodies correlate both with various comorbidities and ADE hallmarks, including increased IL-6 levels and early IgG response. Importantly, anti-Ep9 antibodies can be detected within five days post-symptom onset and sometimes within one day. The results lay the groundwork for a new type of COVID-19 diagnostic for the early prediction of disease severity to guide more effective therapeutic interventions.
Unlocking the potential of personalized medicine in point‐of‐care settings requires a new generation of biomarker and proteomic assays. Ideally, assays could inexpensively perform hundreds of quantitative protein measurements in parallel at the bedsides of patients. This goal greatly exceeds current capabilities. Furthermore, biomarker assays are often challenging to translate from benchtop to clinic due to difficulties achieving and assessing the necessary selectivity, sensitivity, and reproducibility. To address these challenges, we developed an efficient (<5 min), robust (comparatively lower CVs), and inexpensive (decreasing reagent use and cost by >70 %) immunoassay method. Specifically, the immunoblot membrane is dotted with the sample and then developed in a vortex fluidic device (VFD) reactor. All assay steps—blocking, binding, and washing—leverage the unique thin‐film microfluidics of the VFD. The approach can accelerate direct, indirect, and sandwich immunoblot assays. The applications demonstrated include assays relevant to both the laboratory and the clinic.
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