Enzymic hydrolysis of intramolecular complexes for monitoring theophylline in homogeneous competitive protein-binding reactions

Luminescent [6][7][8]. Enzyme channeling immunoassays are applicable to both small and large molecules but have limited sensitivity because of their susceptibility to matrix effects. In these assays, an immune reaction brings two enzymes into proximity on a surface; at that surface, one enzyme produces a product that serves as a chromogenic substrate for the second enzyme [9]. The product reaches higher concentrations near the surface than in the bulk solution, which leads to an increase in the rate of reaction with the proximate second enzyme.We report here on the development of an unusually robust and sensitive homogeneous chemiluminescent immunoassay that is based on a combination of the principles of latex agglutination and channeling. The luminescent oxygen channeling assay (LOCIm'M) principle, which has been described previously [10], uses two different ligand-or receptor-coated polystyrene particles at much lower concentrations than ordinarily used in latex agglutination. ' Typically, particle pairs are formed rather than large aggregates, and photochemically triggered chemiluminescence replaces detection by nephelometry or turbidity. One particle has dissolved in it a photosensitizer that Nonstandard abbreviations: }lBsAg, hepatitis B surfaceantigen;biotin-NIIS ester, biotinyl-6-aminohexanoic acid N-hvdroxvsuccinimide ester; I-lAy, hepatitis A virus;TSH, thyroid-stimulating hormone (thvrotropin);and hCG, human chorionicgonadotropin.

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Enzymic hydrolysis of intramolecular complexes for monitoring theophylline in homogeneous competitive protein-binding reactions

Cited by 5 publications

References 10 publications

Fluorescence and Immunodiagnostic Methods

Fluorescence and Immunodiagnostic Methods

Fluorogenic Substrate Labeled Separation-Free Enzyme Mediated Immunoassays for Haptens and Macromolecules

Homogeneous immunoassays

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