Shekhar Bhansali | PubMed Health

PubMed2007

Sensitive estimation of total cholesterol in blood using Au nanowires based micro-fluidic platform.

Determination of cholesterol level in blood is important in clinical applications. In this work, modified Au nanowires-electrochemical biosensor based on MEMS micro-fluidic platform is proposed for estimating total cholesterol in blood. This sensor consists of "aligned" Au nanowires as working electrode, platinum counter electrode deposited on the silicon platform and Ag/AgCl (3M KCl) reference electrode. The "aligned" Au nanowires are immobilized with cholesterol oxidase and cholesterol esterase using specific covalent chemistry. Further, Au nanowires promotes better electron transfer between the enzymes and electrodes, because of their large surface to volume ratio, small diffusion time, large electrical conductivity and their aligned nature. The modified Au nanowires showed a stable calibration line and a quasi-linear relationship between cholesterol level and current response in the range of 1-6 mM (in steps of 1 mM over the baseline blood serum). The sensitivity of the modified electrode was found to be about 69 nA/mM with good storage and interference stability.

Cholesterol↗

PubMed2006

Ultrasensitive detection of cortisol with enzyme fragment complementation technology using functionalized nanowire.

Cortisol is a member of the glucocorticoid hormone family and a key metabolic regulator. Increased intracellular cortisol levels have been implicated in type 2 diabetes, obesity, and metabolic syndrome. Cortisol is an important bio-marker of stress and its detection is also important in sports medicine. However, rapid methods for sensitive detection of cortisol are limited. Functionalized gold nanowires were used to enhance the sensitivity and selectivity of cortisol detection. Gold nanowires are used to improve the electron transfer between the electrodes. Moreover, the large surface to volume ratio, small diffusion time and high electrical conductivity and their aligned nature will enhance the sensitivity and detection limit of the biosensor several fold. The biosensor was fabricated using, aligned gold (Au) nanowires to behave as the working electrode, platinum deposited on a silicon chip to function as the counter electrode, and silver/silver chloride as reference electrode. The gold nanowires were coupled with cortisol antibodies using covalent linkage chemistry and a fixed amount of 3alpha-hydroxysteroid dehydrogenase was introduced into the reaction cell during each measurement to convert (reduce) ketosteroid into hydroxyl steroid. Furthermore, the micro-fluidic, micro-fluid part of the sensor was fabricated using micro-electro-mechanical system (MEMS) technology to have better control on liquid flow over Au nanowires to minimize the signal to noise ratio. The biosensor was characterized using SEM, AFM and FTIR technique. The response curve of the biosensor was found to be linear in the range of 10-80 microM of cortisol. Moreover, the presence of hydrocortisone is sensitively detected in the range of 5-30 microM. It is concluded that the functionalized gold nanowires with micro-fluidic device using enzyme fragment complementation technology can provide an easy and sensitive assay for cortisol detection in serum and other biological fluids.

3-alpha-Hydroxysteroid Dehydrogenase (B-Specific)↗