Gold bands as a suitable surface for enzyme immunoassays.
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.
Departamento de Química Física y Analítica, Universidad de Oviedo, Oviedo, Asturias, Spain.
Eva M
EM
Abad-Villar
M Teresa
MT
Fernández-Abedul
Agustín
A
Costa-García
D000327:Adsorption; D000469:Alkaline Phosphatase / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D000888:Antibodies, Anti-Idiotypic / Q000032:analysis / Q000276:immunology; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex / Q000032:analysis / Q000276:immunology; D002138:Calibration; D020099:Coated Materials, Biocompatible / Q000138:chemical synthesis; D004800:Enzymes, Immobilized; D004867:Equipment Design; D006046:Gold; D006801:Humans; D007124:Immunoenzyme Techniques / Q000295:instrumentation* / Q000379:methods*; D007075:Immunoglobulin M / Q000032:analysis* / Q000276:immunology; D008422:Materials Testing; D009281:Naphthalenes / Q000737:chemistry; D009943:Organophosphorus Compounds / Q000737:chemistry; D010084:Oxidation-Reduction; D011786:Quality Control; D015203:Reproducibility of Results; D012212:Rheology / Q000295:instrumentation / Q000379:methods; D012680:Sensitivity and Specificity; D013379:Substrate Specificity; D013499:Surface Properties
D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D020099:Coated Materials, Biocompatible; D004800:Enzymes, Immobilized; D007075:Immunoglobulin M; D009281:Naphthalenes; D009943:Organophosphorus Compounds; C044203:anti-IgM; C014694:naphthyl phosphate; D006046:Gold; D000469:Alkaline Phosphatase