Biosensors have recently gained much attention in the field of health care for the management of various important analytes in a biological system. The area achieved tremendous progress from the time when the first Clark electrode for measurement of glucose was realized. Advances in the biosensor design are appearing at a high rate as these devices play increasingly important roles in our daily lives. The increasing incidences of cardiovascular diseases and cardiac arrest are major cause of death of humans world over. One of the most important reasons is hypercholesterolemia, i.e. increased concentration of cholesterol in blood. Hence estimation of cholesterol level in blood is important in clinical applications. This review aims to highlight the recent advances in materials and techniques for cholesterol biosensor design and construction.