Poly(2-thiophene-3-yl-malonic acid) (P3TMA) was synthesized electrochemically using tetrabutylammonium perchlorate (TBAClO(4)) as electrolyte into acetonitrile (ACN) for glucose biosensor application. The properties of P3TMA were investigated using FTIR, SEM, and electrochemical measurements. The glucose oxidase enzyme (GOD) was chemically immobilized onto P3TMA modified electrode using glutaraldehyde as crosslinking. It was observed that response current of P3TMA/GOD enzyme electrode increased linearly with loading glucose concentration. Michaelis Menten constant (K-m) of P3TMA/GOD enzyme electrode was calculated as 0.03517 mM. FTIR and SEM analyses were used to confirm immobilization of GOD onto surface of P3TMA-modified electrode. (C) 2012 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 128: 636-641, 2013