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2.
Colloids Surf B Biointerfaces ; 94: 163-9, 2012 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-22348984

RESUMEN

A procedure for determination of apparent affinity constants K(D)(app) between Concanavalin A (Con A) and naturally d-mannose containing glycoproteins using enzyme-linked lectin assay (ELLA) is reported. Three distinct ELLA protocols are compared to each other with 3 different fitting models used (Liliom, Hill with and without a cooperativity factor). The glycoproteins were physisorbed on a highly charged polystyrene solid surface of immunoassay plates and the amount of lectin bound to the glycoproteins was determined by photometry. The interactions of Con A with five mannose-containing glycoproteins, invertase (INV), glucoamylase (GA), glucose oxidase (GOx), ovalbumin (OVA), and transferrin (TRF) were quantified with apparent affinity constant being in the range 2×10(-7) to 9×10(-6)M. The strength of interaction between Con A and glycoproteins is discussed on the basis of glycan structure/exposure on the protein backbone for each glycoprotein.


Asunto(s)
Concanavalina A/química , Glicoproteínas/química , Inmunoensayo/normas , Sitios de Unión , Glucano 1,4-alfa-Glucosidasa/química , Glucosa Oxidasa/química , Cinética , Manosa/química , Modelos Moleculares , Ovalbúmina/química , Unión Proteica , Transferrina/química , beta-Fructofuranosidasa/química
3.
Biosens Bioelectron ; 13(7-8): 911-23, 1998 Oct 01.
Artículo en Inglés | MEDLINE | ID: mdl-9828388

RESUMEN

Solid binding matrix (SBM) based composite transducers have been used for development of series of multibiosensor systems applicable in various fields. Here we present two hybrid three-channel multibiosensors for simultaneous amperometric operation in food quality control, i.e. glucose/fructose/ethanol multibiosensor, based on glucose oxidase/fructose dehydrogenase/alcohol dehydrogenase surface-modified enzyme electrodes and L-lactate/L-malate/sulfite multibiosensor, based on L-lactate dehydrogenase/L-malate dehydrogenase/sulfite oxidase surface-modified enzyme electrodes. Different parameters have been studied in order to optimize the response of the multibiosensor systems. The multibiosensor showed a good sensitivity, linear range and storage stability. The multibiosensors were used for the determination of glucose, fructose, ethanol, L-lactate, L-malate and sulfite in samples of wine, resulting in a good agreement with data certified by the supplier. Comparison of various designs, surface-modified, bulk-modified and thick-cover, of SBM based biosensors is studied on the example of fructose biosensor.


Asunto(s)
Técnicas Biosensibles , Análisis de los Alimentos , Electroquímica , Etanol/análisis , Fructosa/análisis , Glucosa/análisis , Ácido Láctico/análisis , Malatos/análisis
4.
Anal Chem ; 69(11): 2086-90, 1997 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-21639250

RESUMEN

A new concept of a composite transducer for amperometric biosensors based on the use of a solid substance with amphiphilic character (called a solid binding matrix, SBM) is presented. The electrochemical properties of the transducers prepared with five different SBMs and the characteristics and performance of SBM-based glucose sensors prepared by three different methods are described. Biosensor stability is evaluated and discussed. The biosensor was used for the determination of glucose in wine, yielding results which were consistent with those obtained with the commercially available Glucose Enzyme Photometric Kit. The average accuracy was 6% for the whole range of analyzed concentrations (0.2-47 g/L) using the same sample dilution in a buffer.

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