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Breast cancer is the most common cancer in women worldwide. The detection of biomarkers has played a significant role in the early diagnosis and prognosis of breast cancer. Herein, we describe the construction of a disposable microfluidic immunoarray device (DμID) for the rapid and low-cost detection of CA15-3 (carbohydrate antigen 15-3), a protein biomarker for breast cancer. The DμID was constructed using a simple and rapid prototyping technique and was applied to detect CA15-3 in cancer patients. The DμID construction was based on the use of a double-sided adhesive card with a microfluidic channel and a screen-printed array with 8 electrodes. Both the immunoarray and microfluidic channel were designed using an inexpensive home cutter printer and using low-cost materials. The immunoarray was modified using the layer-by-layer technique aiming at immobilizing the primary antibody. For the biomarker detection, magnetic particles (MPs) modified with polyclonal antibodies and peroxidase enzymes were used as a strategy for capture, separation, and preconcentration of the biomarker, in addition to amplification of the electroanalytical signal. The preconcentration and amplification strategies integrated with the nanostructured immunosensors of the DμID meaningfully contributed toward the detection of CA15-3 with a limit of detection (LoD) of 6 μU mL, requiring as low as 2 μL of serum samples for 8 simultaneous detections. The obtained LoD was 1200 times lower compared to those of other immunosensors previously reported in the literature. The DμID was applied for the detection of CA15-3 in real samples of breast cancer patients and was found to present an excellent correlation with the well-established commercial electrochemiluminescence immunoassay. The association of the DμID with nanostructured surfaces and analyte capturing with bioconjugated paramagnetic particles is essentially a promising breakthrough for the low-cost and accurate detection of cancer biomarkers.

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In a typical protocol for attaching DNA to a gold electrode, thiolated DNA is incubated with the electrode at neutral pH overnight. Here we report fast adsorption of non-thiolated DNA oligomers on gold electrodes at acidic pH (i.e., pH ~3.0). The peak-to-peak potential difference and the redox peak currents in typical cyclic voltammetry of [Fe(CN)6] 3-are investigated to monitor the attachment. Compared with incubation at neutral pH, the lower pH can significantly promote the adsorption processes, enabling efficient adsorption even in 30 min. The adsorption rate is DNA concentration-dependent, while the ionic strength shows no influence. Moreover, the adsorption is base-discriminative, with a preferred order of A >C>>G, T, which is attributed to the protonation of A and C at low pH and their higher binding affinity to gold surface. The immobilized DNA is functional and can hybridize with its complementary DNA but not a random DNA. This work is promising to provide a useful time-saving strategy for DNA assembly on gold electrodes, allowing fast fabrication of DNA-based biosensors and devices.3

Development and application of an electrochemical sensor modified with multi-walled carbon nanotubes and graphene oxide for the sensitive and selective detection of tetracycline

Abstract:With the fast growth of cancer research, new analytical methods are needed to measure anticancer drugs. This is usually accomplished by using sophisticated analytical instruments.Biosensors are attractive candidates for measuring anticancer drugs, but currently few biosensors can achieve this goal. In particular, it is challenging to have a general method to monitor various types of anticancer drugs with different structures. In this work, a biosensor was developed to detect anticancer drugs by modifying carbon paste electrodes with glutathione-s-transferase (GST) enzymes. GST is widely studied in the metabolism of xenobiotics and is a major contributing factor in resistance to anticancer drugs. The measurement of anticancer drugs is based on competition between 1-chloro-2,4-dinitrobenzene (CDNB) and the drugs for the GST enzyme in the electrochemical potential at 0.1 V vs Ag/AgCl by square wave voltammetry (SWV) or using a colorimetric method. The sensor shows a detection limit of 8.8 M cisplatin and exhibits relatively long life time in daily measurements.

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