Cholesterol oxidase (ChOx) has been covalently immobilized onto polyaniline–carboxymethyl cellulose (PANI–CMC) nanocomposite film deposited onto indium-tin-oxide (ITO) coated glass plate using glutaraldehyde as a cross-linker. Fourier transform infrared (FTIR) spectroscopic and electrochemical studies have been used to characterize the PANI–CMC/ITO nanocomposite electrode and ChOx/PANI–CMC/ITO bioelectrode. Scanning electron microscopy (SEM) studies reveal the formation of PANI–CMC nanocomposite fibers of size ∼150 nm in diameter. The ChOx/PANI–CMC/ITO bioelectrode exhibits linearity as 0.5–22 mM, detection limit as 1.31 mM, sensitivity as 0.14 mA/mM cm2, response time as 10 s and shelf-life of about 10 weeks when bioelectrode is stored at 4 °C. The low value of Michaelis-Menten constant (Km) obtained as 2.71 mM reveals high affinity of immobilized ChOx for PANI–CMC/ITO nanocomposite electrode.
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