Fabrication of a label-free electrochemical aptasensor to detect cytochrome c in the early stage of cell apoptosis.

A label-free direct electrochemical aptasensor is presented for the identification of cytochrome c (Cyt c) at the nM concentration level. Carbon nanofibers (CNF), as a highly conductive material, were used to modify a glassy carbon electrode (GCE) and thus increase its conductivity. Moreover, to enhance the immobilization of aptamers (Apt) on the electrode surface, graphene oxide functionalized with aspartic acid (GOAsp) was added to the surface. Aspartic acid with countless carboxyl groups (-COOH) on its surface caused more aptamers to be immobilized on the electrode surface. Electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV), and differential pulse voltammetry (DPV) were used to monitor the step-by-step fabrication of the label-free direct electrochemical aptasensor. The label-free quantification of Cyt c was also done by the direct electron transfer between the Fe(III)/Fe(II)-heme redox-active sites which were selectively bound to the aptamers on the GCE and the surface of the electrode. Under optimum conditions, the peak currents of differential pulse voltammograms at 0.26 V (vs. Ag/AgCl) were used for calibration. The proposed aptasensor performs in a wide dynamic range from 10 nM to 100 µM with a low detection limit of 0.74 nM for cytochrome c. It also has high selectivity as well as acceptable stability. These advantages make the biosensor capable of detecting early-stage apoptotic cells that contribute to early cancer diagnosis.

Monitoring the mechanism of anti-cancer agents to inhibit colorectal cancer cell proliferation: Enzymatic biosensing of glucose combined with molecular docking.

Glucose, a major energy source in cellular metabolism, has a significant role in cell growth. The increase in glucose uptake is a distinguishing hallmark in cancer cells. A key step in glucose utilization is the transport of glucose to the cancer cells for supplying their additional energy. The glucose transporter (or GLUT) family is a membrane protein which facilitates the uptake of glucose in most cancer cell types. Given the increased glucose level in cancer cells and the regulatory role of GLUTs in glucose uptake, it is required to combine both experimental and theoretical studies to develop new methods to monitor cell proliferation. Herein, for the first time, a new strategy was proposed to evaluate the cell proliferation of HT-29 based on glucose consumption in the presence of resveratrol (RSV) as an anticancer agent. A hybrid nanocomposite of carbon nanofibers and nitrogen-doped graphene quantum dots was used to design an enzymatic sensor for the selective and sensitive determination of glucose in cancer cells. The results obtained from the voltammetric technique were compared with the conventional colorimetric assay. A good correlation was observed between the proliferation rate and glucose utilization by cancer cells. As it was observed, RSV induces a decrease in glucose consumption, indicating lower glucose uptake efficiency for HT-29 cells. Molecular docking studies reveal that RSV can block the interaction of glucose with the GLUT family. This is one of the possible mechanisms for the decrease of glucose level followed by the reduction of cell proliferation in the presence of RSV. Compared with traditional methods, in vitro electrochemical techniques benefit from simple, nontoxic, sensitive and low-cost detection assays and hence serve as a novel tool to pursue the growth inhibition of cancer cell in response to anti-cancer agents.

Experimental and theoretical study for miR-155 detection through resveratrol interaction with nucleic acids using magnetic core-shell nanoparticles.

A novel electrochemical nanobiosensor for the detection of miR-155 (as breast cancer biomarker) is introduced . Fe3O4NPs@Ag core-shell nanoparticles were synthesized and their shape and characteristics were confirmed by scanning electron microscope (SEM) imaging, Fourier-transform infrared spectroscopy (FTIR), and X-ray diffraction (XRD) methods. Synthesized nanoparticles were applied onto the magnetic bar carbon paste electrode and then the amine-modified anti-miR-155 (single-stranded probes) was applied on the modified electrode surface and upon hybridization with target miR-155, resveratrol (RSV) was eventually applied as an electrochemical label on the double-strand oligonucleotide. Differential pulse voltammetry (DPV) of the oxidation peak of RSV was assumed as the final signal by sweeping potential from 0 to 0.6 V (vs. Ag/AgCl). The fabrication process was optimized through a series of experiments and the optimized process was confirmed using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The linear range of the fabricated nanobiosensor was 0.5 fM to 1.0 nM and the detection limit was 0.15 fM. The nanobiosensor was able to pass reproducibility and specificity tests using different types of mismatched target sequences.Spiked real samples of human serum were used to confirm that the nanobiosensor enables detection of miR-155 without any significant interferences from other moieties and molecules. Finally, the molecular dynamics simulation of the RSV interaction with single- and double-stranded oligonucleotide was performed and confirmed the preferential binding of RSV to double-stranded DNA; therefore, it can be used as the electrochemical label of DNA and/or miRNA hybridization-based biosensors. Graphical abstract.

Dual-aptamer based electrochemical sandwich biosensor for MCF-7 human breast cancer cells using silver nanoparticle labels and a poly(glutamic acid)/MWNT nanocomposite.

This paper reports on a sensitive and selective method for the detection of Michigan Cancer Foundation-7 (MCF-7) human breast cancer cells and MUC1 biomarker by using an aptamer-based sandwich assay. A biocompatible nanocomposite consisting of multiwall carbon nanotubes (MWCNT) and poly(glutamic acid) is placed on a glassy carbon electrode (GCE). The sandwich assay relies on the use of a mucin 1 (MUC1)-binding aptamer that is first immobilized on the surface of modified GCE. Another aptamer (labeled with silver nanoparticles) is applied for secondary recognition of MCF-7 cells in order to increase selectivity and produce an amplified signal. Differential pulse anodic stripping voltammetry was used to follow the electrochemical signal of the AgNPs. Under the optimal condition, the sensor responds to MCF-7 cells in the concentration range from 1.0 × 102 to 1.0 × 107 cells·mL-1 with a detection limit of 25 cells. We also demonstrate that the MUC1 tumor marker can be detected by the present biosensor. The assay is highly selective and sensitive, acceptably stable and reproducible. This warrants the applicability of the method to early diagnosis of breast cancer. Graphical abstract Schematic of the fabrication of an aptamer-based sandwich biosensor for Michigan Cancer Foundation-7 cells (MCF-7). A MWCNT-poly(glutamic acid) nanocomposite was used as a biocompatible matrix for MUC1-aptamer immobilization. Stripping voltammetry analysis of AgNPs was performed using aptamer conjugated AgNPs as signalling probe.