Gold Nanoparticle-Based Fluorescent Theranostics for Real-Time Image-Guided Assessment of DNA Damage and Repair.

Chemotherapeutic dosing, is largely based on the tolerance levels of toxicity today. Molecular imaging strategies can be leveraged to quantify DNA cytotoxicity and thereby serve as a theranostic tool to improve the efficacy of treatments. Methoxyamine-modified cyanine-7 (Cy7MX) is a molecular probe which binds to apurinic/apyrimidinic (AP)-sites, inhibiting DNA-repair mechanisms implicated by cytotoxic chemotherapies. Herein, we loaded (Cy7MX) onto polyethylene glycol-coated gold nanoparticles (AuNP) to selectively and stably deliver the molecular probe intravenously to tumors. We optimized the properties of Cy7MX-loaded AuNPs using optical spectroscopy and tested the delivery mechanism and binding affinity using the DLD1 colon cancer cell line in vitro. A 10:1 ratio of Cy7MX-AuNPs demonstrated a strong AP site-specific binding and the cumulative release profile demonstrated 97% release within 12 min from a polar to a nonpolar environment. We further demonstrated targeted delivery using imaging and biodistribution studies in vivo in an xenografted mouse model. This work lays a foundation for the development of real-time molecular imaging techniques that are poised to yield quantitative measures of the efficacy and temporal profile of cytotoxic chemotherapies.

Wearable electrochemical glove-based sensor for rapid and on-site detection of fentanyl.

Rapid, on-site detection of fentanyl is of critical importance, as it is an extremely potent synthetic opioid that is prone to abuse. Here we describe a wearable glove-based sensor that can detect fentanyl electrochemically on the fingertips towards decentralized testing for opioids. The glove-based sensor consists of flexible screen-printed carbon electrodes modified with a mixture of multiwalled carbon nanotubes and a room temperature ionic liquid, 4-(3-butyl-1-imidazolio)-1-butanesulfonate). The sensor shows direct oxidation of fentanyl in both liquid and powder forms with a detection limit of 10 µM using square-wave voltammetry. The "Lab-on-a-Glove" sensors, combined with a portable electrochemical analyzer, provide wireless transmission of the measured data to a smartphone or tablet for further analysis. The integrated sampling and sensing methodology on the thumb and index fingers, respectively, enables rapid screening of fentanyl in the presence of a mixture of cutting agents and offers considerable promise for timely point-of-need screening for first responders. Such a glove-based "swipe, scan, sense, and alert" strategy brings chemical analytics directly to the user's fingertips and opens new possibilities for detecting substances of abuse in emergency situations.

Integrating Electrochemical Immunosensing and Cell Adhesion Technologies for Cancer Cell Detection and Enumeration.

We have successfully integrated techniques for controlling cell adhesion and performing electrochemical differential pulse voltammetry (DPV) through the use of digitally controlled microfluidics and patterned transparent indium tin oxide electrode arrays to enable rapid and sensitive enumeration of cancer cells in a scalable microscale format. This integrated approach leverages a dual-working electrode (WE) surface to improve the specificity of the detection system. Here, one of the WE surfaces is functionalized with anti-Melanocortin 1 Receptor antibodies specific to melanoma cancer cells, while the other WE acts as a control (i.e., without antibody), for detecting non-specific interactions between cells and the electrode. The method is described and shown to provide effective detection of melanoma cells at concentrations ranging between 25 to 300 cells per 20 µL sample volume after a 5 min incubation and 15 s of DPV measurements. The estimated limit of detection was ~17 cells. The sensitivity and specificity of the assay were quantified using addition of large fractions of non-target cells and resulted in a detection reproducibility of ~97%. The proposed approach demonstrates a unique integration of electrochemical sensing and microfluidic cell adhesion technologies with multiple advantages such as label-free detection, short detection times, and low sample volumes. Next steps for this platform include testing with patient samples and use of other cell-surface biomarkers for detection and enumeration of circulating tumor cells in prostate, breast, and colon cancer.

Microfluidic-integrated patterned ITO immunosensor for rapid detection of prostate-specific membrane antigen biomarker in prostate cancer.

An optically transparent patterned indium tin oxide (ITO) three-electrode sensor integrated with a microfluidic channel was designed for label-free immunosensing of prostate-specific membrane antigen (PSMA), a prostate cancer (PCa) biomarker, expressed on prostate tissue and circulating tumor cells but also found in serum. The sensor relies on cysteamine capped gold nanoparticles (N-AuNPs) covalently linked with anti-PSMA antibody (Ab) for target specificity. A polydimethylsiloxane (PDMS) microfluidic channel is used to efficiently and reproducibly introduce sample containing soluble proteins/cells to the sensor. The PSMA is detected and quantified by measuring the change in differential pulse voltammetry signal of a redox probe ([Fe(CN)6]3-/[Fe(CN)6]4-) that is altered upon binding of PSMA with PSMA-Ab immobilized on N-AuNPs/ITO. Detection of PSMA expressing cells and soluble PSMA was tested. The limit of detection (LOD) of the sensor for PSMA-based PCa cells is 6/40µL (i.e., 150 cells/mL) (n=3) with a linear range of 15-400 cells/40µL (i.e., 375-10,000 cells/mL), and for the soluble PSMA is 0.499ng/40µL (i.e., 12.5ng/mL) (n=3) with the linear range of 0.75-250ng/40µL (i.e., 19-6250ng/mL), both with an incubation time of 10min. The results indicate that the sensor has a suitable sensitivity and dynamic range for routine detection of PCa circulating tumor cells and can be adapted to detect other biomarkers/cancer cells.

Nanotechnology for Electroanalytical Biosensors of Reactive Oxygen and Nitrogen Species.

Over the past several decades, nanotechnology has contributed to the progress of biomedicine, biomarker discovery, and the development of highly sensitive electroanalytical / electrochemical biosensors for in vitro and in vivo monitoring, and quantification of oxidative and nitrosative stress markers like reactive oxygen species (ROS) and reactive nitrogen species (RNS). A major source of ROS and RNS is oxidative stress in cells, which can cause many human diseases, including cancer. Therefore, the detection of local concentrations of ROS (e. g. superoxide anion radical; O2•- ) and RNS (e. g. nitric oxide radical; NO• and its metabolites) released from biological systems is increasingly important and needs a sophisticated detection strategy to monitor ROS and RNS in vitro and in vivo. In this review, we discuss the nanomaterials-based ROS and RNS biosensors utilizing electrochemical techniques with emphasis on their biomedical applications.

Highly Sensitive Detection and Removal of Lead Ions in Water Using Cysteine-Functionalized Graphene Oxide/Polypyrrole Nanocomposite Film Electrode.

We synthesized cysteine-functionalized graphene oxide (sGO) using carbonyldiimidazole as a cross-linker via amide and carbamate linkages. The sGO/polypyrrole (PPy) nanocomposite film was grown on the working electrode surface of a screen-printed electrode (SPE) via controlled one-step electrochemical deposition. The sGO/PPy-SPE was used to detect lead ions (Pb(2+)) in water by first depositing Pb(2+) on the working electrode surface for 10 min at -1.2 V, and then anodic stripping by differential pulse voltammetry (DPV). The DPV signals were linear in the ranges of 1.4-28 ppb (R(2) = 0.994), 28-280 ppb (R(2) = 0.997), and 280-14 000 ppb (R(2) = 0.990) Pb(2+). The measurable detection limit of the sensor is 0.07 ppb (S/N = 3), which is more than 2 orders of magnitude below the 10 ppb threshold for drinking water set by the World Health Organization. The average removal efficiency of Pb(2+) deposited on the electrode was 99.2% (S/N = 3), with relative standard deviation (RSD) of 3.8%. Our results indicate good affinity of sGO/PPy nanocomposite to Pb(2+), which can be used to effectively adsorb and remove Pb(2+) in water samples. Therefore, sGO/PPy nanocomposite we synthesized is useful for highly sensitive on-site and real-time monitoring of heavy metal ions and water treatment.

An electrochemical immunosensing method for detecting melanoma cells.

An electrochemical immunosensing method was developed to detect melanoma cells based on the affinity between cell surface melanocortin 1 receptor (MC1R) antigen and anti-MC1R antibody (MC1R-Ab). The MC1R-Abs were immobilized in amino-functionalized silica nanoparticles (n-SiNPs)-polypyrrole (PPy) nanocomposite modified on working electrode surface of screen-printed electrode (SPE). Cyclic voltammetry was employed, with the help of redox mediator ([Fe(CN)6](3-)), to measure the change in anodic oxidation peak current arising due to the specific interaction between MC1R antigens and MC1R-Abs when the target melanoma cells are present in the sample. Various factors affecting the sensor performance, such as the amount of MC1R-Abs loaded, incubation time with the target melanoma cells, the presence of interfering non-melanoma cells, were tested and optimized over different expected melanoma cell loads in the range of 50-7500 cells/2.5 mL. The immunosensor is highly sensitive (20 cells/mL), specific, and reproducible, and the antibody-loaded electrode in ready-to-use stage is stable over two weeks. Thus, in conjunction with a microfluidic lab-on-a-chip device our electrochemical immunosensing approach may be suitable for highly sensitive, selective, and rapid detection of circulating tumor cells (CTCs) in blood samples.