Cholesterol biosensor based on cholesterol esterase, cholesterol oxidase and peroxidase immobilized onto conducting polyaniline films

Cholesterol oxidase, cholesterol esterase and peroxidase have been co-immobilized onto electrochemically prepared polyaniline films. These polyaniline–enzyme films characterized using spectroscopic techniques, have been used to fabricate a cholesterol biosensor. This polyaniline-based cholesterol biosensor has a response time of about 240 s, an apparent Km value as 75 mg dl−1 and can be used to estimate cholesterol concentration up to 500 mg dl−1. These polyaniline/cholesterol oxidase/cholesterol esterase films have a detection limit of 25 mg dl−1 with sensitivity of 0.042 μA mg dl−1. The enzyme films were found to be thermally stable up to 48 °C and have a shelf-life of about 6 weeks when stored at 4 °C. The values of the activation energy before and after the critical temperature were found to be 191 and 95.5 kJ mol−1, respectively.

[1]  K. Ramanathan,et al.  Simulation of electrochemical process for glucose oxidase immobilized conducting polymer electrodes , 1996 .

[2]  C. Nicolini,et al.  Electrochemical study of the interaction between cytochrome P450sccK201E and cholesterol. , 2004, Talanta.

[3]  S. Cosnier.  Biomolecule immobilization on electrode surfaces by entrapment or attachment to electrochemically polymerized films. A review. , 1999, Biosensors & bioelectronics.

[4]  J. Zicha,et al.  Abnormalities of membrane function and lipid metabolism in hypertension: a review. , 1999, American journal of hypertension.

[5]  L. Pollegioni,et al.  Cholesterol oxidase from Streptomyces hygroscopicus and Brevibacterium sterolicum: effect of surfactants and organic solvents on activity , 1999, Biotechnology and applied biochemistry.

[6]  J. Kennedy,et al.  Covalent and coordination immobilization of proteins. , 1991, Bioprocess technology.

[7]  Samuel Meites,et al.  Standard Methods of Clinical Chemistry , 1970 .

[8]  N. Murr,et al.  Amperometric determination of cholesterol in serum with use of a renewable surface peroxidase electrode , 1995 .

[9]  G. Wallace,et al.  Conducting polymers and the bioanalytical sciences: new tools for biomolecular communication. A review. , 1996, The Analyst.

[10]  Prem C. Pandey,et al.  Conducting polymer-coated enzyme microsensor for urea , 1988 .

[11]  Martin M. F. Choi,et al.  Hydrogel network entrapping cholesterol oxidase and octadecylsilica for optical biosensing in hydrophobic organic or aqueous micelle solvents. , 2003, Analytical chemistry.

[12]  E. García-Ruiz,et al.  Amperometric cholesterol biosensors based on the electropolymerization of pyrrole and the electrocatalytic effect of Prussian-Blue layers helped with self-assembled monolayers. , 2004, Talanta.

[13]  B. D. Malhotra,et al.  Preparation and characterization of an enzyme electrode based on Cholesterol esterase and Cholesterol oxidase immobilized onto conducting polypyrrole films , 2004 .

[14]  Bansi D. Malhotra,et al.  Immobilization of Lactate Dehydrogenase on Electrochemically Prepared Polyaniline Films , 1999 .

[15]  Bansi D. Malhotra,et al.  Immobilization of lactate dehydrogenase on electrochemically prepared polypyrrole–polyvinylsulphonate composite films for application to lactate biosensors , 2001 .

[16]  B. D. Malhotra,et al.  Coimmobilization of urease and glutamate dehydrogenase in electrochemically prepared polypyrrole-polyvinyl sulfonate films , 2001, Applied biochemistry and biotechnology.

[17]  B. D. Malhotra,et al.  Amperometric cholesterol biosensor based on immobilized cholesterol esterase and cholesterol oxidase on conducting polypyrrole films , 2004 .

[18]  M. Josowicz.  Applications of conducting polymers in potentiometric sensors , 1995 .

[19]  B. D. Malhotra,et al.  Immobilization of lactate dehydrogenase on tetraethylorthosilicate-derived sol-gel films for application to lactate biosensor , 2001, Applied biochemistry and biotechnology.

[20]  Marek Trojanowicz,et al.  Electrochemical biosensors based on enzymes immobilized in electropolymerized films , 1995 .

[21]  Anthony Guiseppi-Elie,et al.  Amperometric determination of cholesterol in serum using a biosensor of cholesterol oxidase contained within a polypyrrole–hydrogel membrane , 2001 .

[22]  Henry Tauber,et al.  Methods of Enzymology. , 1956 .

[23]  Sandro Carrara,et al.  Cholesterol amperometric biosensor based on cytochrome P450scc. , 2004, Biosensors & bioelectronics.

[24]  H. Kramers.  Brownian motion in a field of force and the diffusion model of chemical reactions , 1940 .

[25]  A. Guiseppi-Elie,et al.  Polypyrrole-hydrogel composites for the construction of clinically important biosensors. , 2002, Biosensors & bioelectronics.

[26]  I. Karube,et al.  Amperometric determination of total cholesterol in serum with use of immobilized cholesterol esterase and cholesterol oxidase , 1982 .

[27]  Bansi D. Malhotra,et al.  ENHANCED LOADING OF GLUCOSE OXIDASE ON POLYANILINE FILMS BASED ON ANION EXCHANGE , 1998 .

[28]  J. Cassidy,et al.  Electrochemical Diagnostic Strip Device for Total Cholesterol and Its Subfractions , 2000 .