Polyaniline nanospheres (PANI-NS) prepared by morphological transformation of micelle polymerized camphorsulfonic acid (CSA) doped polyaniline nanotubes (PANI-NT) in the presence of ethylene glycol (EG) have been characterized by X-ray diffraction, atomic force microscopy, transmission electron microscopy, scanning electron microscopy, Fourier transform infra-red and UV-Visible spectroscopy. A PANI-NS (60-80 nm) film deposited onto an indium-tin-oxide (ITO) coated glass plate by the solution casting method has been utilized for covalent immobilization of biomolecules (cholesterol oxidase (ChOx)) viaN-ethyl-N'-(3-dimethylaminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS) chemistry for fabrication of a cholesterol biosensor. The ChOx/PANI-NS/ITO bioelectrode detects cholesterol in the concentration range of 25 to 500 mg dL(-1) with sensitivity of 1.3 x 10(-3) mA mg(-1) dL and regression coefficient of 0.98. Further, this PANI-NS based bioelectrode shows fast response time (10 s), low Michaelis-Menten constant (2.5 mM) and shelf-life of 12 weeks. The spherical nanostructure observed in the final morphology of the PANI-NS film is attributed to hydrogen bonding interactions between PANI-NT and EG.
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