Redox labelled avidin for enzyme sensor architectures.
Conjugates of avidin with ferrocene and with microperoxidase 8 have been used as electrochemically active molecular building blocks. Assemblies of the conjugates with biotinylated glucose oxidase or lactate oxidase on gold electrodes were tested as enzyme sensors for glucose and lactate. The electrochemical detection is based either on ferrocene-mediated oxidation of the substrate in oxygen-free solution, or on microperoxidase-catalysed reduction of H2O2 which is enzymatically produced from the substrate and molecular oxygen. Glucose and lactate were detectable with both detection principles in concentrations down to 1 or 0.1 mM, respectively. The molecular architecture concept allows quick adaptation of the sensors to other analytes, and it provides a platform for arrays of sensors with different selectivity.
Padeste
Celestino
C
Laboratory for Micro- and Nanotechnology, Paul Scherrer Institut, CH-5232 Villigen PSI, Switzerland. celestino.padeste@psi.ch
Steiger
Beat
B
Grubelnik
Andreas
A
Tiefenauer
Louis
L
D001360:Avidin / Q000737:chemistry*; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D020099:Coated Materials, Biocompatible / Q000138:chemical synthesis / Q000737:chemistry; D004800:Enzymes, Immobilized / Q000737:chemistry; D004867:Equipment Design / Q000379:methods; D019544:Equipment Failure Analysis; D005240:Feasibility Studies; D005296:Ferrous Compounds / Q000737:chemistry*; D005947:Glucose / Q000032:analysis* / Q000737:chemistry; D005949:Glucose Oxidase / Q000737:chemistry*; D019344:Lactic Acid / Q000032:analysis* / Q000737:chemistry; D000075163:Metallocenes; D008832:Microchemistry / Q000295:instrumentation / Q000379:methods; D006899:Mixed Function Oxygenases / Q000737:chemistry*; D010084:Oxidation-Reduction; D010544:Peroxidases / Q000737:chemistry*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods
D020099:Coated Materials, Biocompatible; D004800:Enzymes, Immobilized; D005296:Ferrous Compounds; D000075163:Metallocenes; D001360:Avidin; D019344:Lactic Acid; D006899:Mixed Function Oxygenases; D005949:Glucose Oxidase; D010544:Peroxidases; C021799:microperoxidase; C021497:lactate 2-monooxygenase; D005947:Glucose; C004998:ferrocene