Virus Bioresistor (VBR) for Detection of Bladder Cancer Marker DJ-1 in Urine at 10 pM in One Minute

Abstract

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.

Phage-Based Sensors in Medicine: A Review

“…Chemosensors 2020, 8, 61 9 of 26 sensor based on this peptide was developed [65]. The affinity peptides labelled with cysteine were immobilized on the surface of a gold electrode via Au-S self-assembled covalent bond ( Figure 5).…”

“…Chemosensors 2020, 8, 61 11 of 26 charged individuals [80]. Virus bio-resistor is a relatively new approach for sensor construction, which is connected with a stable interest in phage display.…”

“…This comprehensive research allows for the lowering of the LOD to 100 pM (in synthetic urine) and 3.1 nM in PBF (phosphate-buffered fluoride) [56], which is 2 or 3 orders of magnitude lower than that reported for the previous VBR biosensor [82]. Since initial descriptions of M13 bacteriophage embedded into the PEDOT ultra-thin layer, several sensing [56,59,61,[80][81][82][83] devices were constructed and examined. First, a VBR point-ofproof biosensor was constructed using a modified PSMA-binding peptide reported previously (LDCVEVFQNSCDW) [55].…”

“…Additionally, a thorough analysis of the theoretical aspects of virus bioresistors was presented. In a result, LOD was reduced to 7.5 nM for HSA detection with a response time of 3-30 s. Recently, a novel VBR sensor for detection of DJ-1 protein-a marker of urinary bladder cancer has been presented [61]. The DJ-1-binding peptide was initially established to be KYRYVCHDVGGTYCIRD*V, where (*) means ATG codon (STOP codon-termination of protein Recently, a novel VBR sensor for detection of DJ-1 protein-a marker of urinary bladder cancer has been presented [61].…”

“…In a result, LOD was reduced to 7.5 nM for HSA detection with a response time of 3-30 s. Recently, a novel VBR sensor for detection of DJ-1 protein-a marker of urinary bladder cancer has been presented [61]. The DJ-1-binding peptide was initially established to be KYRYVCHDVGGTYCIRD*V, where (*) means ATG codon (STOP codon-termination of protein Recently, a novel VBR sensor for detection of DJ-1 protein-a marker of urinary bladder cancer has been presented [61]. The DJ-1-binding peptide was initially established to be KYRYVCHDVGGTYCIRD*V, where (*) means ATG codon (STOP codon-termination of protein biosynthesis.…”