title: Glutaraldehyde cross-linked glutamate oxidase coated microelectrode arrays: selectivity and resting levels of glutamate in the CNS.
abstract: Glutaraldehyde is widely used as a cross-linking agent for enzyme immobilization onto microelectrodes. Recent studies and prior reports indicate changes in enzyme activity and selectivity with certain glutaraldehyde cross-linking procedures that may jeopardize the performance of microelectrode recordings and lead to falsely elevated responses in biological systems. In this study, the sensitivity of glutaraldehyde cross-linked glutamate oxidase-based microelectrode arrays to 22 amino acids was tested and compared to glutamate. As expected, responses to electroactive amino acids (Cys, Tyr, Trp) were detected at both nonenzyme-coated and enzyme-coated microelectrodes sites, while the remaining amino acids yielded no detectable responses. Electroactive amino acids were effectively blocked with a m-phenylene diamine (mPD) layer and, subsequently, no responses were detected. Preliminary results on the use of poly(ethylene glycol) diglycidyl ether (PEGDE) as a potentially more reliable cross-linking agent for the immobilization of glutamate oxidase onto ceramic-based microelectrode arrays are reported and show no significant advantages over glutaraldehyde as we observe comparable selectivities and responses. These results support that glutaraldehyde-cross-linked glutamate oxidase retains sufficient enzyme specificity for accurate in vivo brain measures of tonic and phasic glutamate levels when immobilized using specific "wet" coating procedures.
authors: Department of Anatomy and Neurobiology, Parkinson's Disease Translational Research Center of Excellence, University of Kentucky, Lexington, KY 40536-0098, United States.
authors: Jason J
authors: JJ
authors: Burmeister
authors: Verda A
authors: VA
authors: Davis
authors: Jorge E
authors: JE
authors: Quintero
authors: Francois
authors: F
authors: Pomerleau
authors: Peter
authors: P
authors: Huettl
authors: Greg A
authors: GA
authors: Gerhardt
mesh_terms: D000594:Amino Acid Oxidoreductases / Q000187:drug effects* / Q000502:physiology; D015374:Biosensing Techniques; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D004800:Enzymes, Immobilized / Q000187:drug effects* / Q000502:physiology; D018698:Glutamic Acid / Q000032:analysis*; D005976:Glutaral / Q000494:pharmacology*; D008839:Microelectrodes
chemical_list: D003432:Cross-Linking Reagents; D004800:Enzymes, Immobilized; D018698:Glutamic Acid; D000594:Amino Acid Oxidoreductases; C023061:L-glutamate oxidase; D005976:Glutaral
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