Cholesterol amperometric biosensor based on cytochrome P450scc.
A screen-printed enzyme electrode based on flavocytochrome P450scc (RfP450scc) for amperometric determination of cholesterol has been developed. A one-step method for RfP450scc immobilization in the presence of glutaraldehyde or by entrapment of enzyme within a hydrogel of agarose is discussed. The sensitivity of the biosensor based on immobilization procedures of flavocytochrome P450scc by glutaric aldehyde is 13.8 nA microM(-1) and the detection limit is 300 microM with a coefficient of linearity 0.98 for cholesterol in the presence of sodium cholate as detergent. The detection limits and the sensitivity of the agarose-based electrode are 155 microM and 6.9 nA microM(-1) with a linearity coefficient of 0.99. For both types of electrodes, the amperometric response to cholesterol in the presence of detergent was rather quick (1.5-2 min).
Shumyantseva
Victoria
V
Institute of Biomedical Chemistry, Pogodinskaya St., 10, Moscow, 119121 Russia. victoria@ibmh.msk.su
Deluca
Gilda
G
Bulko
Tatiana
T
Carrara
Sandro
S
Nicolini
Claudio
C
Usanov
Sergei A
SA
Archakov
Alexander
A
D015374:Biosensing Techniques; D002784:Cholesterol / Q000032:analysis*; D002786:Cholesterol Side-Chain Cleavage Enzyme; D004566:Electrodes; D006108:Graphite; D012238:Rhodium; D012834:Silver; D018030:Silver Compounds
D018030:Silver Compounds; D012834:Silver; D006108:Graphite; D002784:Cholesterol; D012238:Rhodium; D002786:Cholesterol Side-Chain Cleavage Enzyme; C037548:silver chloride