TY - JOUR
T1 - Analysis of NAD(P)+ and NAD(P)H cofactors by means of imprinted polymers associated with Au surfaces
T2 - A surface plasmon resonance study
AU - Raitman, Oleg A.
AU - Chegel, Vladimir I.
AU - Kharitonov, Andrei B.
AU - Zayats, Maya
AU - Katz, Eugenii
AU - Willner, Itamar
PY - 2004/2/16
Y1 - 2004/2/16
N2 - Crosslinked films consisting of the acrylamide-acrylamidophenylboronic acid copolymer that are imprinted with recognition sites for β-nicotinamide adenine dinucleotide (NAD+), β-nicotinamide adenine dinucleotide phosphate NADP+, and their reduced forms (NAD(P)H), are assembled on Au-coated glass supports. The binding of the oxidized cofactors NAD+ or NADP+ or the reduced cofactors NADH or NADPH to the respective imprinted sites results in the swelling of the polymer films through the uptake of water. Surface plasmon resonance (SPR) spectroscopy is employed to follow the binding of the different cofactors to the respective imprinted sites. The imprinted recognition sites reveal selectivity towards the association of the imprinted cofactors. The method enables the analysis of the NAD(P)+ and NAD(P)H cofactors in the concentration range of 1×10-6 to 1×10-3M. The cofactor-imprinted films associated with the Au-coated glass supports act as active interfaces for the characterization of biocatalyzed transformations that involve the cofactor-dependent enzymes. This is exemplified with the characterization of the biocatalyzed oxidation of lactate to pyruvate in the presence of NAD+ and lactate dehydrogenase using the NADH-imprinted polymer film.
AB - Crosslinked films consisting of the acrylamide-acrylamidophenylboronic acid copolymer that are imprinted with recognition sites for β-nicotinamide adenine dinucleotide (NAD+), β-nicotinamide adenine dinucleotide phosphate NADP+, and their reduced forms (NAD(P)H), are assembled on Au-coated glass supports. The binding of the oxidized cofactors NAD+ or NADP+ or the reduced cofactors NADH or NADPH to the respective imprinted sites results in the swelling of the polymer films through the uptake of water. Surface plasmon resonance (SPR) spectroscopy is employed to follow the binding of the different cofactors to the respective imprinted sites. The imprinted recognition sites reveal selectivity towards the association of the imprinted cofactors. The method enables the analysis of the NAD(P)+ and NAD(P)H cofactors in the concentration range of 1×10-6 to 1×10-3M. The cofactor-imprinted films associated with the Au-coated glass supports act as active interfaces for the characterization of biocatalyzed transformations that involve the cofactor-dependent enzymes. This is exemplified with the characterization of the biocatalyzed oxidation of lactate to pyruvate in the presence of NAD+ and lactate dehydrogenase using the NADH-imprinted polymer film.
KW - Imprinted polymers
KW - NAD(P) and NAD(P)H cofactors
KW - Sensor
KW - Surface plasmon resonance
UR - http://www.scopus.com/inward/record.url?scp=1642575133&partnerID=8YFLogxK
U2 - 10.1016/S0003-2670(03)00511-7
DO - 10.1016/S0003-2670(03)00511-7
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AN - SCOPUS:1642575133
SN - 0003-2670
VL - 504
SP - 101
EP - 111
JO - Analytica Chimica Acta
JF - Analytica Chimica Acta
IS - 1
ER -