ABSTRACT
Separative extended gate field effect transistor (SEGFET) type devices have been used as an ion sensor or biosensor as an alternative to traditional ion sensitive field effect transistors (ISFETs) due to their robustness, ease of fabrication, low cost and possibility of FET isolation from the chemical environment. The layer-by-layer technique allows the combination of different materials with suitable properties for enzyme immobilization on simple platforms such as the extended gate of SEGFET devices enabling the fabrication of biosensors. Here, glucose biosensors based on dendrimers and metallophthalocyanines (MPcs) in the form of layer-by-layer (LbL) films, assembled on indium tin oxide (ITO) as separative extended gate material, has been produced. NH(3)(+) groups in the dendrimer allow electrostatic interactions or covalent bonds with the enzyme (glucose oxidase). Relevant parameters such as optimum pH, buffer concentration and presence of serum bovine albumin (BSA) in the immobilization process were analyzed. The relationship between the output voltage and glucose concentration shows that upon detection of a specific analyte, the sub-products of the enzymatic reaction change the pH locally, affecting the output signal of the FET transducer. In addition, dendritic layers offer a nanoporous environment, which may be permeable to H(+) ions, improving the sensibility as modified electrodes for glucose biosensing.
Subject(s)
Biosensing Techniques/instrumentation , Dendrimers/chemistry , Glucose/analysis , Indoles/chemistry , Nickel/chemistry , Transistors, Electronic , Aspergillus niger/enzymology , Aziridines/chemistry , Buffers , Glucose Oxidase/metabolism , Hydrogen-Ion Concentration , Isoindoles , RecyclingABSTRACT
The effect of gamma-radiation doses of 12.5-380 kGy on the infrared spectra, gel content, mechanical properties, and the release of oxobutyl-5-fluoro-2'-deoxyuridine (OfdUrd, an antitumor agent) from poly(ethylene-co-vinyl acetate) (EVA) films was studied. The results showed that the application of radiation doses produced a crosslinking reaction leading to a maximum gel content of about 85% in the case of 150 kGy. Higher doses did not increase the gel content in EVA films. The mechanical properties (tensile strength, percentage elongation at break and Young's modulus) of all studied EVA matrices were affected by the exposure to gamma-radiation. Irradiation doses over 50 kGy caused an increase in the Young's modulus of EVA and at the same time a decrease in the strain per cent. Moreover, the network structure formed after irradiation reduced significantly the OFdUrd release from EVA films. In this manner, the radiation dose applied to the polymeric matrix modulated the release of OFdUrd, avoiding the high concentrations that may cause severe systemic toxicity. The loading of OFdUrd to EVA film triggered a slight hyperemia after implantation, while the inflammatory reaction was only observed during the first two days.
Subject(s)
Delayed-Action Preparations/chemistry , Drug Carriers/chemistry , Membranes, Artificial , Polyvinyls/chemistry , Uridine/analogs & derivatives , Animals , Cross-Linking Reagents/chemistry , Cross-Linking Reagents/radiation effects , Diffusion , Elasticity/radiation effects , Gamma Rays , Materials Testing , Polyvinyls/radiation effects , Rats , Rats, Wistar , Uridine/administration & dosage , Uridine/chemistryABSTRACT
A method has been developed in which a layer of p-aminosalicylic acid (4-amino-2-hydroxybenzoic acid) (PAS), a water soluble pharmaceutical compound of the nonsteroidal anti-inflammatory drug (NSAID) class with antiaggregant platelet activity, is covalently immobilized onto a segmented polyurethane, Biospan (SPU) surface. Thus, SPU surfaces were modified by grafting of hexamethylenediisocyanate. and the free isocyanate remaining on the SPU surface were then coupled through a condensation reaction to amine groups of p-aminosalicylic acid. The bonding of PAS from aqueous solution onto SPU surface was studied by ATR-FTIR. UV and fluorescence spectroscopy. Plateau levels of coupled PAS were reached within 1.2 microg/cm2 using PAS solution concentrations of 1mg/ ml. The surface wettability of the polymeric films measured by contact angle indicate that the introduction of the PAS turns the surface more hydrophilic (theta(water) = 43.1 +/- 2.1) relatively to the original SPU films (theta(water) = 70.3 +/- 1.9). The in vitro albumin (BSA) adsorption shows that the PAS-SPU films adsorb more BSA (250/microgmm2) than the original SPU (112 microg mm2). Thrombogenicity was assessed by measuring the thrombus formation and platelet adhesion of the SPU containing PAS relatively to nonmodified SPU surfaces. The polymeric surfaces with immobilized PAS had better nonthrombogenic characteristics as indicated by the low platelet adhesion, high adsorption of albumin relatively to fibrinogen and low thrombus formation, making them potentially good candidates for biomedical applications.
Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Biocompatible Materials , Polyurethanes/chemistry , Albumins/chemistry , Aminosalicylic Acid/chemistry , Animals , Blood Platelets/cytology , Cattle , Cell Adhesion , Cyanates/chemistry , Factor XII/metabolism , Humans , Kinetics , Models, Chemical , Protein Binding , Spectrometry, Fluorescence , Time Factors , Ultraviolet Rays , Water/chemistryABSTRACT
This paper reports on the use of the crude extract of avocado (CEA) fruit (Persea americana) as a source of tyrosinase enzyme. CEA was immobilized via layer by layer (LbL) technique onto indium tin oxide (ITO) substrates and applied in the detection of monophenol using a potentiometric biosensor. Poly(propylene imine) dendrimer of generation 3 (PPI-G3) was used as a counter ion in the layer by layer process due to its highly porous structure and functional groups suitable for enzyme linkage. After the immobilization of the crude CEA as multilayered films, standard samples of monophenol were detected in the 0.25-4.00 mM linear range with approximately 28 mV mM(-1) of sensitivity. This sensitivity is 14 times higher than the values found in the literature for a similar system. The results show that it is possible to obtain efficient and low-cost biosensors for monophenol detection using potentiometric transducers and alternative sources of enzymes without purification.