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We present a novel and efficient nanoporous microfluidic biochip consisting of a functionalized chitosan/anatase titanium dioxide nanoparticles (antTiO2-CH) electrode integrated in a polydimethylsiloxane (PDMS) microchannel assembly. The electrode surface can be enzyme functionalized depending on the application. We studied in detail cholesterol sensing using the cholesterol esterase (ChEt) and cholesterol oxidase (ChOx) functionalized chitosan supported mesoporous antTiO2-CH microfluidic electrode. The available functional groups present in the nanoporous antTiO2-CH surface in this microfluidic biochip can play an important role for enzyme functionalization, which has been quantified by the X-ray photoelectron spectroscopic technique. The Brunauer-Emmett-Teller (BET) studies are used to quantify the specific surface area and nanopore size distribution of titania nanoparticles with and without chitosan. Point defects in antTiO2 can increase the heterogeneous electron transfer constant between the electrode and enzyme active sites, resulting in improved electrochemical behaviour of the microfluidic biochip. The impedimetric response of the nanoporous microfluidic biochip (ChEt-ChOx/antTiO2-CH) shows a high sensitivity of 6.77 kΩ (mg dl(-1))(-1) in the range of 2-500 mg dl(-1), a low detection limit of 0.2 mg dl(-1), a low Michaelis-Menten constant of 1.3 mg dl(-1) and a high selectivity. This impedimetric microsystem has enormous potential for clinical diagnostics applications.

We report the fabrication of an efficient, label-free, selective and highly reproducible immunosensor with unprecedented sensitivity (femto-molar) to detect a breast cancer biomarker for early diagnostics. Mesoporous zinc oxide nanofibers (ZnOnFs) are synthesized by electrospinning technique with a fiber diameter in the range of 50-150 nm. Fragments of ZnOnFs are electrophoretically deposited on an indium tin oxide glass substrate and conjugated via covalent or electrostatic interactions with a biomarker (anti-ErbB2; epidermal growth factor receptor 2). Oxygen plasma treatment of the carbon doped ZnOnFs generates functional groups (-COOH, -OH, etc.) that are effective for the conjugation of anti-ErbB2. ZnOnFs without plasma treatment that conjugate via electrostatic interactions were also tested for comparison. Label-free detection of the breast cancer biomarker by this point-of-care device is achieved by an electrochemical impedance technique that has high sensitivity (7.76 kΩ μM(-1)) and can detect 1 fM (4.34 × 10(-5) ng mL(-1)) concentration. The excellent impedimetric response of this immunosensor provides a fast detection (128 s) in a wide detection test range (1.0 fM-0.5 μM). The oxy-plasma treated ZnOnF immunoelectrode shows a higher association constant (404.8 kM(-1) s(-1)) indicating a higher affinity towards the ErbB2 antigen compared to the untreated ZnOnF immunoelectrode (165.6 kM(-1) s(-1)). This sensor is about an order of magnitude more sensitive than the best demonstrated in the literature based on different nanomaterials and about three orders of magnitude better than the ELISA standard for breast cancer biomarker detection. This proposed point-of-care cancer diagnostic offers several advantages, such as higher stability, rapid monitoring, simplicity, cost-effectiveness, etc., and should prove to be useful for the detection of other bio- and cancer markers.

Urease (Urs) and glutamate dehydrogenase (GLDH) have been covalently co-immobilized onto a self-assembled monolayer (SAM) comprising of 10-carboxy-1-decanthiol (CDT) via EDC-NHS chemistry deposited onto one of the two patterned gold (Au) electrodes for estimation of urea using poly(dimethylsiloxane) based microfluidic channels (2 cm × 200 μm × 200 μm). The CDT/Au and Urs-GLDH/CDT/Au electrodes have been characterized using Fourier transform infrared (FTIR) spectroscopy, contact angle (CA), atomic force microscopy (AFM) and electrochemical cyclic voltammetry (CV) techniques. The electrochemical response measurement of a Urs-GLDH/CDT/Au bioelectrode obtained as a function of urea concentration using CV yield linearity as 10 to 100 mg dl(-1), detection limit as 9 mg dl(-1) and high sensitivity as 7.5 μA mM(-1) cm(-2).