A sensitive sandwich enzyme immunoassay for calcitonin gene-related peptide (CGRP): characterization and application.
Thirty mouse monoclonal antibodies (mAbs) directed against rat calcitonin gene-related peptide-alpha (CGRP-alpha) have been obtained. These mAbs are classified in 2 groups, one recognizing the peptide N-terminus and the other binding the C-terminus. A two-site immunometric assay was developed using mAb CGRP-83 as capture antibody, whereas mAb CGRP-72 acts as tracer, covalently labeled with enzyme acetylcholinesterase. This assay appeared sensitive (limit of detection: 2 pg/ml) and precise, allowing quantitative measurement of all human and murine CGRP isoforms. The assay was used to determine specific concentrations of CGRP in different rat, mice and guinea pig samples. The validity of the test was demonstrated by HPLC fractionation experiments.
Frobert
Y
Y
CEA, Service de Pharmacologie et d'Immunologie, DRM, CEA-Saclay, Gif s/Yvette, France.
Nevers
M C
MC
Amadesi
S
S
Volland
H
H
Brune
P
P
Geppetti
P
P
Grassi
J
J
Créminon
C
C
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D015740:Calcitonin Gene-Related Peptide / Q000097:blood / Q000302:isolation & purification* / Q000378:metabolism; D002211:Capsaicin / Q000494:pharmacology; D002490:Central Nervous System / Q000737:chemistry / Q000187:drug effects; D000939:Epitopes; D006168:Guinea Pigs; D006801:Humans; D007124:Immunoenzyme Techniques / Q000379:methods*; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D020033:Protein Isoforms / Q000302:isolation & purification; D051381:Rats; D017207:Rats, Sprague-Dawley; D012137:Respiratory System / Q000737:chemistry / Q000187:drug effects; D014018:Tissue Distribution
D000911:Antibodies, Monoclonal; D000939:Epitopes; D020033:Protein Isoforms; D015740:Calcitonin Gene-Related Peptide; D002211:Capsaicin