Surface functionalization plays a crucial role in the design of biosensors. For this purpose, a novel functional monomer, 6-(4,7-bis(2,3-dihydrothieno[3,4-b][1,4]dioxin-5-yl)-2H-benzo[d][1,2,3]triazol-2-yl)hexan-1-amine (BEDOA-6), was designed and synthesized. Poly(BEDOA-6) was utilized as an immobilization matrix for glucose oxidase biosensor construction. Moreover, polymethylmethacrylate (PMMA) layered silicate nanocomposites were prepared by in situ suspension polymerization. Conducting polymer surface was modified with PMMA/clay nanocomposite material and a glucose biosensor was developed. In addition, XPS and SEM were utilized to characterize the surface properties. The biosensor shows a wide linear range between 2.8 mu M and 1.2mM to glucose with a low detection limit of 1.99 mu M. Finally, the biosensor was tested on serum samples containing actual human blood. The results were in well-agreement with a reference method.