This work, describes for the first time, a simple biosensing design to yield an ultrasensitive electrochemical biosensor for a cancer biomarker detection, miRNA-155, with linear response down to the attomolar range. MiRNA-155 was selected for being overexpressed in breast cancer. The biosensor was assembled in two stages: (1) the immobilization of the anti-miRNA-155 that was thiol modified on an Au-screen printed electrode (Au-SPE), followed by (2) blocking the areas of non-specific binding with mercaptosuccinic acid. Atomic force microscopy (AFM) and electrochemical techniques including cyclic voltammetry (CV), impedance spectroscopy (EIS) and square wave voltammetry (SWV) confirmed the surface modification of these devices and their ability to hybridize successfully and stably with miRNA-155. The final biosensor provided a sensitive detection of miRNA-155 from 10 aM to 1.0 nM with a low detection limit (LOD) of 5.7 aM in real human serum samples. Good results were obtained in terms of selectivity towards breast cancer antigen CA-15.3 and bovine serum albumin (BSA). Raw fluid extracts from cell-lines of melanoma did not affect the biosensor response (no significant change of the blank), while raw extracts from breast cancer yielded a positive signal against miRNA-155. This simple and sensitive strategy is a promising alternative for simultaneous quantitative analysis of multiple miRNA in physiological fluids for biomedical research and point-of-care (POC) diagnosis.
1 February 2016 in Sensors and Actuators B: Chemical
This research work presents, for the first time, a screen-printed electrode (SPE) made on a PCB board with silver tracks (Ag) and a three electrode configuration (AgxO-working, AgxO-counter and Ag/AgxO-reference electrodes), following the same approach as printed-circuit boards (PCBs). This low cost and disposable device was tested for screening a cancer biomarker in point-of-care. The selected biomarker was carcinogenic embryonic antigen (CEA) protein, routinely used to follow-up the progression of specific cancer diseases.
Serious brain disorders, such as the Alzheimer's Disease (AD), are associated with a marked drop in the levels of important neurotransmitters, such as acetylcholine (ACh). Real time monitoring of such biomarkers can therefore play a critical role in enhancing AD therapies by allowing timely diagnosis, verifications of treatment effectiveness, and developments of new medicines. In this study, we present the first acetylcholine/oxygen hybrid enzymatic fuel cell for the self-powered on site detection of ACh in plasma, which is based on the combination of an enzymatic anode with a Pt cathode. Firstly, an effective acetylcholinesterase immobilized electrode was developed and its electrochemical performance evaluated. Highly porous gold was used as the electrode material, and the enzyme was immobilized via a one step rapid and simple procedure that does not require the use of harsh chemicals or any electrode/enzyme pre-treatments. The resulting enzymatic electrode was subsequently used as the anode of a miniature flow-through membrane-less fuel cell and showed excellent response to varying concentrations of ACh. The peak power generated by the fuel cell was 4nW at a voltage of 260mV and with a current density of 9μAcm-2. The limit of detection of the fuel cell sensor was 10μM, with an average response time as short as 3min. These exciting results open new horizons for point-of-care Alzheimer diagnosis and provide an attractive potential alternative to established methods that require laborious and time-consuming sample treatments and expensive instruments.