Marisa Maltez-da Costa

Rapid identification and quantification of tumor cells using an electrocatalytic method based on gold nanoparticles.

There is a high demand for simple, rapid, efficient, and user-friendly alternative methods for the detection of cells in general and, in particular, for the detection of cancer cells. A biosensor able to detect cells would be an all-in-one dream device for such applications. The successful integration of nanoparticles into cell detection assays could allow for the development of this novel class of cell sensors. Indeed, their application could well have a great future in diagnostics, as well as other fields. As an example of a novel biosensor, we report here an electrocatalytic device for the specific identification of tumor cells that quantifies gold nanoparticles (AuNPs) coupled with an electrotransducing platform/sensor. Proliferation and adherence of tumor cells are achieved on the electrotransducer/detector, which consists of a mass-produced screen-printed carbon electrode (SPCE). In situ identification/quantification of tumor cells is achieved with a detection limit of 4000 cells per 700 microL of suspension. This novel and selective cell-sensing device is based on the reaction of cell surface proteins with specific antibodies conjugated with AuNPs. Final detection requires only a couple of minutes, taking advantage of the catalytic properties of AuNPs on hydrogen evolution. The proposed detection method does not require the chemical agents used in most existing assays for the detection of AuNPs. It allows for the miniaturization of the system and is much cheaper than other expensive and sophisticated methods used for tumor cell detection. We envisage that this device could operate in a simple way as an immunosensor or DNA sensor. Moreover, it could be used, even by inexperienced staff, for the detection of protein molecules or DNA strands.

Electrochemical detection of Salmonella using gold nanoparticles.

A disposable immunosensor for Salmonella enterica subsp. enterica serovar Typhimurium LT2 (S) detection using a magneto-immunoassay and gold nanoparticles (AuNPs) as label for electrochemical detection is developed. The immunosensor is based on the use of a screen-printed carbon electrode (SPCE) that incorporates a permanent magnet underneath. Salmonella containing samples (i.e. skimmed milk) have been tested by using anti-Salmonella magnetic beads (MBs-pSAb) as capture phase and sandwiching afterwards with AuNPs modified antibodies (sSAb-AuNPs) detected using differential pulse voltammetry (DPV). A detection limit of 143 cells mL(-1) and a linear range from 10(3) to 10(6) cells mL(-1) of Salmonella was obtained, with a coefficient of variation of about 2.4%. Recoveries of the sensor by spiking skimmed milk with different quantities of Salmonella of about 83% and 94% for 1.5×10(3) and 1.5×10(5) cells mL(-1) were obtained, respectively. This AuNPs detection technology combined with magnetic field application reports a limit of detection lower than the conventional commercial method carried out for comparison purposes in skimmed milk samples.

Simple monitoring of cancer cells using nanoparticles.

Here we present a new strategy for a simple and fast detection of cancer circulating cells (CTCs) using nanoparticles. The human colon adenocarcinoma cell line (Caco2) was chosen as a model CTC. Similarly to other adenocarcinomas, colon adenocarcinoma cells have a strong expression of EpCAM, and for this reason this glycoprotein was used as the capture target. We combine the capturing capability of anti-EpCAM functionalized magnetic beads (MBs) and the specific labeling through antibody-modified gold nanoparticles (AuNPs), with the sensitivity of the AuNPs-electrocatalyzed hydrogen evolution reaction (HER) detection technique. The fully optimized process was used for the electrochemical detection of Caco2 cells in the presence of monocytes (THP-1), other circulating cells that could interfere in real blood samples. Therefore we obtained a novel and simple in situ-like sensing format that we applied for the rapid quantification of AuNPs-labeled CTCs in the presence of other human cells.

Controlling the electrochemical deposition of silver onto gold nanoparticles: Reducing interferences and increasing the sensitivity of magnetoimmuno assays

An electrocatalytical method induced by gold nanoparticles in order to improve the sensitivity of the magnetoimmunosensing technology is reported. Microparamagnetic beads as primary antibodies immobilization platforms and gold nanoparticles modified with secondary antibodies as high sensitive electrocatalytical labels are used. A built-in magnet carbon electrode allows the collection/immobilization on its surface of the microparamagnetic beads with the immunological sandwich and gold nanoparticle catalysts attached onto. The developed magnetoimmunosensing technology allows the antigen detection with an enhanced sensitivity due to the catalytic effect of gold nanoparticles on the electroreduction of silver ions. The main parameters that affect the different steps of the developed assay are optimized so as to reach a high sensitive electrochemical detection of the protein. The low levels of gold nanoparticles detected with this method allow the obtaining of a novel immunosensor with low protein detection limits (up to 23 fg/mL), with special interest for further applications in clinical analysis, food quality and safety as well as other industrial applications.

Gold nanoparticle-based electrochemical magnetoimmunosensor for rapid detection of anti-hepatitis B virus antibodies in human serum

A sandwich immunoassay using magnetic beads as bioreaction platforms and AuNPs as electroactive labels for the electrochemical detection of human IgG antibodies anti-Hepatitis B surface antigen (HBsAg), is here presented as an alternative to the standard methods used in hospitals for the detection of human antibodies directed against HBsAg (such as ELISA or MEIA). The electrochemical detection of AuNPs is carried out approaching their catalytic properties towards the hydrogen evolution in an acidic medium, without previous nanoparticle dissolution. The obtained results are a good promise toward the development of a fully integrated biosensing set-up. The developed technology based on this detection mode would be simple to use, low cost and integrated into a portable instrumentation that may allow its application even at doctor-office. The sample volumes required can be lower than those used in the traditional methods. This may lead to several other applications with interest for clinical control.

Detection of Circulating Cancer Cells Using Electrocatalytic Gold Nanoparticles

A rapid cancer cell detection and quantification assay, based on the electrocatalytic properties of gold nanoparticles towards the hydrogen evolution reaction, is described. The selective labeling of cancer cells is performed in suspension, allowing a fast interaction between the gold nanoparticle labels and the target proteins expressed at the cell membrane. The subsequent electrochemical detection is accomplished with small volumes of sample and user-friendly equipment through a simple electrochemical method that generates a fast electrochemical response used for the quantification of nanoparticle-labeled cancer cells. The system establishes a selective cell-detection assay capable of detecting 4 × 10(3) cancer cells in suspension that can be extended to several other cells detection scenarios.