Nylon-6,6, enzyme immobilization

Figure 2 Effect of enzyme immobilization on luminescent image spatial resolution evaluated using coupled enzymatic reactions on nylon net as a model system, (a) Immobilized 3a-hydroxysteroid dehydrogenase (b) immobilized 3a-hydroxysteroid dehydrogenase and FMN-NADH oxidoreductase (c) immobilized 3a-hydroxysteroid dehydrogenase, FMN-NADH oxidoreductase, and bacterial luciferase. (From Ref. 47. Copyright John Wiley Sons Ltd. Reproduced with permission.)...

Immobilized enzymes may be used in affinity chromatographic methods but their use as catalysts may be in either the production or removal of compounds in chemical processes or as analytical tools. Many substrate assays can be performed using enzymes immobilized on a variety of surfaces, e.g. glass beads, plastic or nylon tubing. Alternatively they may be incorporated into gel or microparticulate layers on dry strips or slides. 

In the previous papers(12,13), we reported on the vessel access type, i.e. tubular type, glucose sensor. It consisted of a glucose electrode system with a GOX enzyme immobilized Nylon membrane and a glucose semipermeable membrane, and a reference oxygen electrode system. The sensor could directly measure up to 700 mg/dl of BGL in an arterial blood stream when it was placed into an external A-V shunt. This sensor, however, has some problems such as thrombus during in vivo testing without heparin and clinical complexity associated with implanting the sensor in a blood stream. 

FEP Teflon FEP membrane PMSP Poly(1-trimethylsilyl-l-propyne) membrane Nz Enzyme immobilized Nylon filter... 

Calibration profiles of the sensor based on the final nylon-enzyme net (III) were disappointing oonpared with the analogous sensor based on nylon net type II. The lower detection limit was only 0.1 mM glucose and currents produced were about 80% smaller. However, this alternative immobilization scheme serves to illustrate the synthetic versatility of nylon-6,6 in the biosensor field. 

Choline oxidase and acetylcholineesterase Enzymes immobilized on a nylon net attached to H202-selective amperometric sensor. ChO is used for choline and AChE and ChO for acetylcholine. Rectilinear response in the range of 1-10 pM. Response time 1-2 min. Interferences occur from ascorbic acid, primary amines, and most seriously from betaine aldehyde. [64]... 

Two main criteria for the membrane selection are pore size and material. As peroxidases usually have sizes in the range of 10-80 kDa, ultrafiltration membranes with a molecular cutoff between 1 and 50 kDa are the most adequate to prevent enzyme leakage [99]. The materials commonly applied to ultrafiltration membranes are synthetic polymers (nylon, polypropylene, polyamide, polysulfone, cellulose and ceramic materials [101]. The adequate material depends on a great number of variables. When enzyme is immobilized into the matrix, this must be prepared at mild conditions to preserve the enzymatic activity. In the case of enzyme immobilization onto the membrane, this should be activated with the reactive groups necessary to interact with the functional groups of the enzyme. If an extractive system is considered, the selection of the hydrophilicity or hydro-phobicity of the membrane should be performed according to the features of reactants, products, and solvents. In any case, the membrane should not interfere with the catalytic integrity of the enzyme. 

Matrices for enzyme immobilization on nylon tubes, nylon-covered glass and packed reactors have been investigated in detail [369]. The results have shown that nylon-supported enzymes are less active but easier to use than those supported on glass whiskers . 

Enzymes can be immobilized by matrix entrapment, by microencapsulation, by physical or ionic adsorption, by covalent binding to organic or inorganic polymer-carriers, or by whole cell immobilization (5 ). Particularly impressive is the great number of chemical reactions developed for the covalent binding of enzymes to inorganic carriers such as glass, to natural polymers such as cellulose or Sepharose, and to synthetic polymers such as nylon, polyacrylamide, and other vinyl polymers and... 

Among potentiometric enzyme sensors, the urea enzyme electrode is the oldest (and the most important). The original version consisted of an enzyme layer immobilized in a polyacrylamide hydrophilic gel and fixed in a nylon netting attached to a Beckman 39137 glass electrode, sensitive to the alkali metal and NHj ions [19, 2A Because of the poor selectivity of this glass electrode, later versions contained a nonactin electrode [20,22] (cf. p. 187) and especially an ammonia gas probe [25] (cf. p. 72). This type of urea electrode is suitable for the determination of urea in blood and serum, at concentrations from 5 to 0.05 mM. Figure 8.2 shows the dependence of the electrode response... 

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