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Construction and application of the amperometric uric acid biosensors based on the covalent immobilization of uricase by different strategies
Tvorynska, Sofiia ; Barek, J. ; Josypčuk, Bohdan
In this work, a promising combination of a biosensor based on the\nenzymatic mini-reactor with the detection principle of four-electron\nreduction of the consumed oxygen at highly negative potential has\nbeen developed for uric acid determination using flow injection\nanalysis. The construction of the biosensor provides a spatial\nsegregation of the biorecognition (uricase-based mini-reactor) and\ndetection (tubular detector of silver solid amalgam (TD-p-AgSA))\nparts. To find out the most appropriate enzyme immobilization\nprotocol, three different strategies of the covalent attachment for\nuricase from Bacillus fastidiosus have been compared. It was found\nthat the biosensor with the mini-reactor based on the covalent\nattachment of uricase via glutaraldehyde to -NH2 functionalized\nmesoporous silica powder MCM-41 showed extremely high stability\n(>1 year) and reusability (at least 600 measurements) The biosensor's\npractical applicability was confirmed by successful determination\nof uric acid in human urine.
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