A sensitive sandwich enzyme immunoassay for calcitonin gene-related peptide (CGRP): Characterization and application

Authors
Frobert, Y Nevers, MC Amadesi, S Volland, H Brune, P Geppetti, P Grassi, J Creminon, C
Citation
Y. Frobert et al., A sensitive sandwich enzyme immunoassay for calcitonin gene-related peptide (CGRP): Characterization and application, PEPTIDES, 20(2), 1999, pp. 275-284
Citations number
37
Categorie Soggetti
Biochemistry & Biophysics
Journal title
PEPTIDES
ISSN journal
01969781 → ACNP
Volume
20
Issue
2
Year of publication
1999
Pages
275 - 284
Database
ISI
SICI code
0196-9781(1999)20:2<275:ASSEIF>2.0.ZU;2-J
Abstract
Thirty mouse monoclonal antibodies (mAbs) directed against rat calcitonin g ene-related peptide-oc (CGRP-cr) have been obtained. These mAbs are classif ied in 2 groups, one recognizing the peptide N-terminus and the other bindi ng the C-terminus. A two-site immunometric assay was developed using mAb CG RP-83 as capture antibody, whereas mAb CGRP-72 acts as tracer, covalently l abeled with enzyme acetylcholinesterase. This assay appeared sensitive (lim it of detection: 2 pg/ml) and precise, allowing quantitative measurement of all human and murine CGRP isoforms. The assay was used to determine specif ic concentrations of CGRP in different rat, mice and guinea pig samples. Th e validity of the test was demonstrated by HPLC fractionation experiments. (C) 1999 Elsevier Science Inc. All rights reserved.