Abstract
Gold bands sputtered over a polymeric material, Kapton, are employed for the development of enzyme immunoassays. The immunological interaction takes place between human IgM and alkaline phosphatase (AP) conjugated anti-IgM. The model analyte (IgM) could be determined following a non-competitive design in the range of 0.05-5 ppm, with a limit of detection of 50 ppb. After the interaction, gold bands are sequentially inserted in a flow system and the extension of the reaction is followed through the enzymatic hydrolysis of naphthylphosphate, AP substrate. The product, naphthol, is oxidised to naphtoquinone in the gold band of the flow cell that constitutes the detector. Parameters affecting the interaction are studied and calibration curves are performed. The reproducibility between different bands (RSD=4%, n=5) and possibilities of regeneration are also detailed.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adsorption
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Alkaline Phosphatase / analysis*
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Alkaline Phosphatase / chemistry
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Alkaline Phosphatase / immunology
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Antibodies, Anti-Idiotypic / analysis
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Antibodies, Anti-Idiotypic / immunology
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Antibodies, Monoclonal
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Antigen-Antibody Complex / analysis
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Antigen-Antibody Complex / immunology
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Calibration
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Coated Materials, Biocompatible* / chemical synthesis
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Enzymes, Immobilized
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Equipment Design
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Gold*
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Humans
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Immunoenzyme Techniques / instrumentation*
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Immunoenzyme Techniques / methods*
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Immunoglobulin M / analysis*
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Immunoglobulin M / immunology
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Materials Testing*
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Naphthalenes / chemistry
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Organophosphorus Compounds / chemistry
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Oxidation-Reduction
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Quality Control
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Reproducibility of Results
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Rheology / instrumentation
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Rheology / methods
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Sensitivity and Specificity
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Substrate Specificity
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Surface Properties
Substances
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Antibodies, Anti-Idiotypic
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Antibodies, Monoclonal
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Antigen-Antibody Complex
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Coated Materials, Biocompatible
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Enzymes, Immobilized
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Immunoglobulin M
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Naphthalenes
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Organophosphorus Compounds
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anti-IgM
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naphthyl phosphate
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Gold
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Alkaline Phosphatase