title: 3-(4-Hydroxyphenyl)propionic acid: the forgotten detection substrate for ligand-binding assay-based bioanalysis.
abstract: 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.
authors.affiliation: Roche Pharma Research & Early Development, Pharmaceutical Sciences, Global DMPK & Bioanalytical R&D, Roche Innovation Center Munich, Germany.
authors.forename: Gregor
authors.initials: G
authors.lastname: Jordan
authors.affiliation: Roche Pharma Research & Early Development, Pharmaceutical Sciences, Global DMPK & Bioanalytical R&D, Roche Innovation Center Munich, Germany.
authors.forename: Kay-Gunnar
authors.initials: KG
authors.lastname: Stubenrauch
authors.affiliation: Roche Pharma Research & Early Development, Pharmaceutical Sciences, Global DMPK & Bioanalytical R&D, Roche Innovation Center Munich, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Heinrich
authors.affiliation: Roche Pharma Research & Early Development, Pharmaceutical Sciences, Global DMPK & Bioanalytical R&D, Roche Innovation Center Munich, Germany.
authors.forename: Roland F
authors.initials: RF
authors.lastname: Staack
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid; D003124:Colorimetry; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006735:Horseradish Peroxidase / Q000378:metabolism; D006736:Horses; D007118:Immunoassay; D007074:Immunoglobulin G / Q000737:chemistry*; D008024:Ligands; D008252:Macaca fascicularis; D008297:Male; D013058:Mass Spectrometry; D051379:Mice; D010636:Phenols / Q000097:blood*; D011422:Propionates / Q000097:blood*; D011786:Quality Control; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: C491266:3-hydroxyphenylpropionic acid; D007074:Immunoglobulin G; D008024:Ligands; D010636:Phenols; D011422:Propionates; D006735:Horseradish Peroxidase
keywords: 3-(4-hydroxyphenyl)propionic acid; HPPA; LC–MS; dynamic range; fluorogenic substrate; ligand-binding assay; sensitivity