title: Covalent enzyme immobilization by poly(ethylene glycol) diglycidyl ether (PEGDE) for microelectrode biosensor preparation.
abstract: Poly(ethylene glycol) diglycidyl ether (PEGDE) is widely used as an additive for cross-linking polymers bearing amine, hydroxyl, or carboxyl groups. However, the idea of using PEGDE alone for immobilizing proteins on biosensors has never been thoroughly explored. We report the successful fabrication of microelectrode biosensors based on glucose oxidase, d-amino acid oxidase, and glutamate oxidase immobilized using PEGDE. We found that biosensors made with PEGDE exhibited high sensitivity and a response time on the order of seconds, which is sufficient for observing biological processes in vivo. The enzymatic activity on these biosensors was highly stable over several months when they were stored at 4 °C, and over at least 3d at 37 °C. Glucose microelectrode biosensors implanted in the central nervous system of anesthetized rats reliably monitored changes in brain glucose levels induced by sequential administration of insulin and glucose. PEGDE provides a simple, low cost, non-toxic alternative for the preparation of in vivo microelectrode biosensors.
authors.affiliation: INSERM U1028, CNRS UMR5292, Lyon Neuroscience Research Center, Integrated Physiology of the Waking State, Lyon, F-69000, France.
authors.forename: Natalia
authors.initials: N
authors.lastname: Vasylieva
authors.forename: Bogdan
authors.initials: B
authors.lastname: Barnych
authors.forename: Anne
authors.initials: A
authors.lastname: Meiller
authors.forename: Caroline
authors.initials: C
authors.lastname: Maucler
authors.forename: Loredano
authors.initials: L
authors.lastname: Pollegioni
authors.forename: Jian-Sheng
authors.initials: JS
authors.lastname: Lin
authors.forename: Daniel
authors.initials: D
authors.lastname: Barbier
authors.forename: Stéphane
authors.initials: S
authors.lastname: Marinesco
mesh_terms: D000594:Amino Acid Oxidoreductases; D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D001923:Brain Chemistry; D003432:Cross-Linking Reagents; D003605:D-Amino-Acid Oxidase; D004800:Enzymes, Immobilized; D004853:Epoxy Resins; D005404:Fixatives; D005947:Glucose / Q000032:analysis; D005949:Glucose Oxidase; D018698:Glutamic Acid / Q000032:analysis; D005976:Glutaral; D008297:Male; D051381:Rats; D017208:Rats, Wistar; D012694:Serine / Q000032:analysis
chemical_list: D003432:Cross-Linking Reagents; D004800:Enzymes, Immobilized; D004853:Epoxy Resins; D005404:Fixatives; C035364:Quetol 651; D018698:Glutamic Acid; D012694:Serine; D005949:Glucose Oxidase; D000594:Amino Acid Oxidoreductases; C023061:L-glutamate oxidase; D003605:D-Amino-Acid Oxidase; D005947:Glucose; D005976:Glutaral