A biosensor is an integrated bioanalytical device that employs biological elements as an analyte sensor which coupled to a transducer for signal detection. Biosensor provides sensitive and selective determination of various compounds of analytical relevance. Recently, use of nanomaterials is gaining attention as biosensor matrix. Nanomaterials provide biocompatible environment to immobilize biomolecules and facilitate electron transfer for biosensor fabrication. Enzyme electrodes based on glucose oxidase (GOx)have played a leading role in the move to simple easy-to-use blood sugar testing and are expected to play a similar role in going forward continuous glucose monitoring. The present book highlights the preparation, characterization and application of nanomaterials and their composites for development of glucose biosensor. The fabricated bioelectrodes were tested for various parameters like pH, temperature, interferents etc and have been utilized for glucose estimation. The results indicate that the nanomaterials and their composites provide useful matrices for the fabrication of commercial glucose sensing bioelectrodes.