Enzymes attachment, cellulose acetate

The enzyme is used in an enzyme electrode in which a tube is sealed at its lower end with a cellulose acetate membrane. An outer membrane of collagen is also attached to the end of the electrode tube and glucose oxidase enzyme is contained in the space between the two diaphragms. 

Enzymes have been attached to a nylon matrix (135-13 7), a pig intestine (138), the hydrophobic membrane of a gas-selective sensor (139), and controlled pore glass (140). Recent comparative studies of the coupling agents GA and benzoquinone support the preferential use of benzoquinone for binding GOD to nylon mesh (13 7), or to cellulose acetate membranes (141) with lysine (13 7). Both investigations report robust electrode behaviors with respect to prolonged exposure to glucose, while lifetimes of the membrane electrodes were ca. 3 months. 

The enzyme ascorbate oxidase (AOD) is immobilized over Clark-type oxygen electrodes. The biocatalyst is attached to polyamide nets with GA (252), cross-linked with collagen-glutaraldehyde (251) or albumin-GA (253), or linked to cellulose acetate membranes (253). 

A three-enzyme electrode system, such as needed for creatinine measurement, poses a more difficult enzyme-immobilisation problem, in that different enzymes have different immobilisation requirements and their microenvironmental interrelationships need to be optimised. For one creatine sensor, the requisite creatine amidinohydrolase and sarcosine oxidase were immobihsed in polyurethane pre-polymer and PEG-hnked creatinine amidohydrolase was attached via diisocyanate pre-polymer to create a polyurethane adduct [14]. The likelihood of enzyme inactivation with chemical immobih-sation is high, but provided an enzyme preparation survives this, long-term stability is feasible. In the case of these three particular enzymes, a loss of activity resulted from silver ions diffusing from the reference electrode the material solution was to protect the enzyme layer with a diffusion-resisting cellulose acetate membrane. 

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