Lin, Zhihong and Wu, Meng and Wolfbeis, Otto S. and Schaeferling, Michael (2006) A novel method for time-resolved fluorimetric determination and imaging of the activity of peroxidase, and its application to an enzyme-linked immunosorbent assay. CHEMISTRY-A EUROPEAN JOURNAL, 12 (10). pp. 2730-2738. ISSN 0947-6539,
Full text not available from this repository. (Request a copy)Abstract
A new type of fluorescence assay for the determination of peroxidase (POx) activity is presented. The assay is based on the indication of the enzymatic consumption of H2O2 (HP), using a fluorescent europium-tetracycline (Eu3TC) complex as indicator. On addition of HP, this complex forms a highly fluorescent adduct (Eu3TC-HP), which is decomposed in the presence of POx to form the weakly fluorescent europium-tetracycline (EU3TC). Hence, the activity of the enzyme can be directly determined by means of the luminescent EU3TC complex as indicator. The POx assay demonstrated herein was elaborated starting from a spectral characterization of the complex systems involved. Due to the long lifetime of lanthanide luminescence, both steady-state and time-resolved luminescence assays can easily be performed. The time-resolved assay can quantify POx in the range from 4.0x10(-5) to 5.9x10(-3) U mL(-1), with a limit of detection of 1.0x10(-5) U mL(-1). The effects of POx inhibitors such as cyanide, hydroxylamine, and azide have also been studied. In addition, a time-resolved fluorescent detection method for a POx-based enzyme-linked immunosorbent assay (ELISA) has been developed, which is demonstrated in a sandwich model assay with bovine IgG serving as analyte. Furthermore, a time-resolved fluorescent imaging method is demonstrated that makes use of a straightforward imaging set-up adjusted to the optical properties of the europium reagent.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | AMPLIFIED LANTHANIDE LUMINESCENCE; HORSERADISH-PEROXIDASE; HYDROGEN-PEROXIDE; OXIDATIVE ENZYMES; SUBSTRATE-BINDING; GLUCOSE-OXIDASE; EUROPIUM PROBE; FLUORESCENCE; COMPLEX; ACID; fluorescent probes; imaging agents; immunoassays; peroxidase activity; time-resolved spectrometry |
| Subjects: | 500 Science > 540 Chemistry & allied sciences |
| Divisions: | Chemistry and Pharmacy > Institut für Analytische Chemie, Chemo- und Biosensorik > Chemo- und Biosensorik (Prof. Antje J. Bäumner, formerly Prof. Wolfbeis) |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 17 Feb 2021 12:28 |
| Last Modified: | 17 Feb 2021 12:28 |
| URI: | https://pred.uni-regensburg.de/id/eprint/34780 |
Actions (login required)
 |
View Item |
