In the present study, highly sensitive and high-throughput optical waveguidebiosensors were fabricated by using the sensing membranes containing dye andpolymer-enzyme complex. Optical light waveguide can detect the optical change inthe vicinity of the guide surface with high sensitivity due to the evanescentwave scattering. The glucose sensing membranes, composed of dye, enzymes, andbiocompatible polymers were prepared by solution processing on the opticalwaveguide. Herein, we used 3, 3’, 5,5’-tetramethylbenzidine (TMBZ) as a dye, glucose oxidase (GOD) andperoxidase (POD) as enzymes, phosphatide polymer for protection of biologicalactivity of enzyme, and carboxymethyl cellulose (CMC) as a binder. Then wefocused on the optimal composition and structure of sensing membranes for theenhancement in the sensitivity and response speed. The developed glucose sensorsdemonstrated 20 times higher sensitivity than the conventional light waveguideglucose sensors and the low-detection limit of 0.1g/L glucose within thedetection time of 60 sec. For further improvement in the sensitivity,microporous sensing membranes were fabricated by using electrosprayingtechnique. The electroprayed sensing membranes gave 40 % higher sensitivity thannonporous sensing membranes. These results show that both the composition andstructure of sensing membrane are crucial factors for highly sensitive andhigh-throughput optical waveguide biosensors.