title: Glucose level determination with a multi-enzymatic cascade reaction in a functionalized glass chip.
abstract: In this work we show the functionalization of the interior of microfluidic glass chips with poly(2-hydroxyethyl methacrylate) polymer brushes as anchors for co-immobilization of the enzymes glucose-oxidase and horseradish peroxidase. The formation of the brush layer and subsequent immobilization of these enzymes have been characterized on flat surfaces by atomic force microscopy and Fourier transform infrared spectroscopy, and studied inside glass chips by field emission scanning microscopy. Enzyme-functionalized glass chips have been applied for performing a multi-enzymatic cascade reaction for the fast (20 s) determination of glucose in human blood samples and the result is in excellent agreement with values obtained from the conventional hospital laboratory. The limit of detection of this bi-enzymatic method is 60 μM. With the advantages of high selectivity and reproducibility, this functionalization method can be used for improving the efficiency of glucose sensors.
authors.affiliation: Dipartimento di Chimica, Sapienza Università di Roma, p.le Aldo Moro 5, 00185, Rome, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Costantini
authors.forename: Roald
authors.initials: R
authors.lastname: Tiggelaar
authors.forename: Simona
authors.initials: S
authors.lastname: Sennato
authors.forename: Francesco
authors.initials: F
authors.lastname: Mura
authors.forename: Stefan
authors.initials: S
authors.lastname: Schlautmann
authors.forename: Federico
authors.initials: F
authors.lastname: Bordi
authors.forename: Han
authors.initials: H
authors.lastname: Gardeniers
authors.forename: Cesare
authors.initials: C
authors.lastname: Manetti
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D005898:Glass / Q000737:chemistry*; D005947:Glucose / Q000032:analysis* / Q000737:chemistry*; D005949:Glucose Oxidase / Q000737:chemistry / Q000378:metabolism*; D006735:Horseradish Peroxidase / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D010084:Oxidation-Reduction; D012825:Silicon / Q000737:chemistry; D013056:Spectrophotometry, Ultraviolet; D013499:Surface Properties
chemical_list: D004800:Enzymes, Immobilized; D006861:Hydrogen Peroxide; D005949:Glucose Oxidase; D006735:Horseradish Peroxidase; D005947:Glucose; D012825:Silicon