A novel type of working electrode based on MWCNT is fabricated on silicon substrate for detection of total cholesterol. Multiwall carbon nanotubes (MWNT), which are vertically grown on a silicon platform, promote heterogeneous electron transfer between the enzyme and the working electrode. Immobilization of cholesterol oxidase (Pseudomonas fluorescens.) on the electrode is done by entrapment of the enzyme in a bio-friendly, water soluble polymer, polyvinyl alcohol (PVA), which also increases the hydrophilicity of the carbon nanotube (CNT) surface (with experimental evidence) and thereby facilitating the intimate attachment of the protein molecules. Cyclic voltammograms exhibit efficient detection of cholesterol in the range of 100-300 mg/dl, which covers the entire range of possible cholesterol concentration in human blood. The influence of interferents, like glucose, ascorbic acid and uric acid, present in the physiological fluids, is also examined. Owing to its compatibility with standard silicon microfabrication technology, the signal processing circuitry can also be integrated onto the same silicon substrate yielding a compact miniaturized biochip.