A sensitive and specific two-site, sandwich-amplified enzyme immunoassay for neuropeptide Y.
The development of a new enzyme immunoassay for neuropeptide Y (NPY) is reported. Two monoclonal antibodies directed against distinct epitopes of NPY are used, one as a capture antibody (NPY02) and the other one as an indicator antibody (NPY05), this latter antibody being labeled with alkaline phosphatase. The assay calibration curve was performed over concentrations of 1 to 250 pM in a NPY-free plasma. The intra-assay coefficient of variation (CV) ranged from 0.025 to 11.9%, whereas the interassay CV was comprised between 5 and 12%. The limit of detection of this assay was 1 pM (100 amol/well). Neuropeptide Y levels are related to sampling conditions; basal concentrations of NPY with low SEM are found when less than 1.2 ml of blood is taken in EDTA tubes, the sample is centrifuged at 4 degrees C, and immediately frozen. Unanesthetized spontaneously hypertensive rats exhibited higher NPY plasma concentrations than normotensive Wistar-Kyoto controls (53 +/- 7 pM and 25 +/- 2 pM, respectively, mean +/- SEM, p < 0.01). Plasma NPY levels are similar in 16- and 36-week-old animals. In conclusion, this technique makes it possible to assay a large number of samples within 24 h without requiring radioactivity.
Grouzmann
E
E
Hypertension Division, University Hospital, Lausanne, Switzerland.
Aubert
J F
JF
Waeber
B
B
Brunner
H R
HR
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D006973:Hypertension / Q000097:blood; D007124:Immunoenzyme Techniques; D008969:Molecular Sequence Data; D009478:Neuropeptide Y / Q000097:blood*; D051381:Rats; D011918:Rats, Inbred SHR; D011921:Rats, Inbred WKY; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D009478:Neuropeptide Y