[9] Immobilization of enzymes on nylon

Hornby, W.E.; Goldstein, Léon

doi:10.1016/s0076-6879(76)44011-9

Cited by 53 publications

(19 citation statements)

References 28 publications

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“…For activation of the released amino groups the nonwoven was activated by 15-min treatment with 20 mL of a 5% (w/v) solution of glutaraldehyde in 0.2 M boric acid/sodium borate (pH 8.5) at 20°C (Hornby and Goldstein, 1976;Onyezili, 1987). Nonreacted glutaraldehyde was removed by washing with 3 × 20 mL of 0.1 M potassium phosphate (pH 7.5).…”

Section: Activation Of Polyamidementioning

confidence: 99%

“…But, until now, the covalent immobilization of enzymes to polyamide nonwovens has been employed only rarely, although some useful methods have been reported (Hornby and Goldstein, 1976;Morris et al, 1975). Two severe restrictions hamper the extensive preparation of enzyme-immobilized polyamide surfaces: (1) the absence of strongly reactive groups; and (2) unfavorable interactions of the enzyme with the weakly polar surface (Lozano and Iborra, 1997).…”

Section: Introductionmentioning

confidence: 99%

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Immobilization of thermolysin to polyamide nonwoven materials

Moeschel

Nouaimi

Steinbrenner

et al. 2003

Biotech & Bioengineering

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In the last few years, an increasing number of biotechnological techniques have been applied to the restoration and conservation of works of art, paintings, old maps, and papers or books. Enzymes can solve problems that give restorers difficulties, although for many applications it is not possible to use soluble enzymes; therefore, it is necessary to look for suitable carriers for immobilization. Different methods for covalent immobilization of enzymes to polyamide nonwovens were tested, using thermolysin as an example. Two distinct strategies were pursued: (1). controlled, partial hydrolysis of the polymer and subsequent binding of the enzyme to the released amino and carboxy groups; and (2). attachment of reactive groups directly to the polyamide without disintegrating the polymeric structure (O-alkylation). Different spacers were used for covalent fixation of the enzyme in both cases. The enzyme was fixed to the released amino groups by glutaraldehyde, either with or without a spacer. Either way, active enzyme could be immobilized to the matrix. However, intense treatment caused severe damage to the stability of the nonwoven fabric, and reduced the mechanical strength. Conditions were investigated to conserve the nonwoven fabric structure while obtaining near-maximum immobilized enzyme activity. Immobilization of the enzyme to the released carboxy group after acid hydrolysis was performed using dicyclohexylcarbodiimide. In comparison to the enzyme bound via the amino group, the yield of immobilized enzyme activity was slightly lower when benzidine was taken as spacer and still lower with a 1,6-hexanediamine spacer. O-alkylation performed with dimethylsulfate caused severe damage to the nonwoven fabric structure. Considerably better results were obtained with triethyloxonium tetrafluoroborate. As the spacers 1,6-hexanediamine and adipic acid dihydrazide were used, activation for immobilizing thermolysin was performed with glutaraldehyde, adipimidate, and azide. With the exception of azide, all combinations of spacers and activation reagents gave high yields of immobilized enzyme activity. Thermolysin immobilized by this technique showed a remarkably improved stability with respect to elevated temperature, extreme pH values, and reduced polarity. The nonwoven fabric can be stored for weeks without loss of enzyme activity by washing with distilled water and drying.

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Section: Activation Of Polyamidementioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Immobilization of thermolysin to polyamide nonwoven materials

Moeschel

Nouaimi

Steinbrenner

et al. 2003

Biotech & Bioengineering

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“…As noted in several reviews, chemiluminescence-based detection methods have a number of attractive features including: [29][30][31] ease and rapidity of the assay, ultra-high sensitivity, high signal stability (steady glow of light), wide linear range (over several orders of magnitude), and relatively simple instrumentation (requires only a light detector apparatus). For our model bioconjugation studies, nylon 6 was chosen as an enzyme support material because it is inexpensive and non-biodegradable, [32,33] which renders possible prolonged exposure to biological media without impairment of its structural integrity. In addition, nylon-based membrane filter supports have previously been shown to be particularly effective enhancers of the chemiluminescent signal intensity in alkaline phosphatase/1,2-dioxetane-based systems when compared against other nonnylon-based supports (e.g., nitrocellulose membranes) that are also commonly used in conventional solid-phase nucleic acid detection assays.…”

Section: Introductionmentioning

confidence: 99%

Bioconjugation of Alkaline Phosphatase to Mechanically Processed, Aqueous Suspendible Electrospun Polymer Nanofibers for Use in Chemiluminescent Detection Assays

Mark

Stolper

Baratti

et al. 2008

Macromolecular Bioscience

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Aqueous suspendible polymer nanostructures were prepared by simple microtome processing of electrospun nylon 6 nanofibers and were used to immobilize calf intestinal alkaline phosphatase (ALP) by either covalent or noncovalent bioconjugation chemistries. It was found that noncovalent immobilization of ALP to the mechanically cut nanofibers (mean length approximately 4 microm; mean diameter approximately 80 nm) using a multi-stacked, layer-by-layer (LBL) approach with the cationic polymer Sapphire II resulted in the highest enzyme loading (48.1 +/- 0.4 microg . mg(-1) nanofiber) when compared to other covalent immobilization methods based on glutaraldehyde crosslinking. The biofunctionalized nanofibers were also characterized for their chemiluminescent activity with the dioxetane substrate, CSPD. The results indicate that the kinetic parameters, K(m) and V(max), for the catalytic activity of the nanostructure-bound ALP enzyme were influenced by the particular types of immobilization methods employed. In terms of the overall catalytic performance of the various immobilized ALP systems, a single-stacked LBL assembly approach resulted in the highest level of enzymatic activity per unit mass of nanofiber support. To the best of our knowledge, this study represents the first report examining the preparation of mechanically shortened, aqueous dispersed electrospun polymer nanofibers for potential application as enzyme scaffolds in chemiluminescent-based assay systems.

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“…These ap- [1][2][3] proaches confer to the membrane hydrophilic properties. An alternative way to activate the membrane structure is grafting by means of ␥-radiation.…”

Section: Introductionmentioning

confidence: 99%

Immobilization of ?-galactosidase on nylon membranes grafted with diethylenglycol dimethacrylate (DGDA) by ?-radiation: Effect of membrane pore size

Eldin

Maio

Martino

et al. 1999

Adv. Polym. Technol.

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The dependence of grafting and catalytic activity on the pore size of nylon membranes grafted with different DGDA concentrations and loaded with β‐galactosidase is discussed. Membrane pore sizes were 0.2, 1.2, and 3.0 μm. Grafting and enzyme activity, all other experimental conditions being the same, decrease with the increase of membrane pore size. The membranes have been characterized also with reference to pH and temperature. The experimental conditions giving the best performance for each of the three membranes have been identified. In all cases the membranes giving the best results are those having 0.2 μm pore size. The advantages of using in non‐isothermal bioreactors a single membrane, catalytic and hydrophobic at the same time, are also indicated. © 1999 John Wiley & Sons, Inc. Adv Polym Techn 18: 109–123, 1999

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[9] Immobilization of enzymes on nylon

Cited by 53 publications

References 28 publications

Immobilization of thermolysin to polyamide nonwoven materials

Immobilization of thermolysin to polyamide nonwoven materials

Bioconjugation of Alkaline Phosphatase to Mechanically Processed, Aqueous Suspendible Electrospun Polymer Nanofibers for Use in Chemiluminescent Detection Assays

Immobilization of ?-galactosidase on nylon membranes grafted with diethylenglycol dimethacrylate (DGDA) by ?-radiation: Effect of membrane pore size

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