Journal
BIOANALYSIS
Volume 9, Issue 4, Pages 407-418Publisher
FUTURE SCI LTD
DOI: 10.4155/bio-2016-0225
Keywords
3-(4-hydroxyphenyl)propionic acid; dynamic range; fluorogenic substrate; HPPA; LC-MS; ligand-binding assay; sensitivity
Funding
- Roche Diagnostics GmbH
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Ligand-binding assays are ideal for routine bioanalysis, but we reason that the straightforward replacement of the conventional chromogenic horseradish peroxidase substrate, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid, of a routinely used preclinical immunoassay to detect hIgG, with the fluorogenic 3-(4-hydroxyphenyl) propionic acid would broaden the narrow dynamic range. The replacement leads to a sensitivity of 0.47 (minimum required dilution [MRD] 10) and 1.02 (MRD 50) ng/ml, and dynamic ranges of 3.3 (MRD 10) and 3.6 (MRD 50) orders of magnitude, and thereby had improved sensitivity and dynamic range compared with other conventional colorimetric ELISAs, other ligand-binding assay technologies or LC-MS assays. Improvements in sensitivity and dynamic range were achieved for the sera of horse, mice and monkeys without assay optimization.
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