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Clinical Chemistry 31: 861-863, 1985;
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Clinical Chemistry, Vol 31, 861-863, Copyright © 1985 by American Association for Clinical Chemistry

Development and evaluation of a radioimmunoassay for Arg8-vasopressin, after extraction with Sep-Pak C18

KA Ysewijn-Van Brussel and AP De Leenheer

We describe a specific double-antibody radioimmunoassay for measuring arginine vasopressin (AVP) in human plasma. Antisera of high avidity were obtained from rabbits that had been injected with AVP coupled to bovine thyroglobulin. The antibody reacts with both the tripeptide tail and the pentapeptide ring of the molecule, thereby eliminating cross reaction with oxytocin. Synthetic AVP was labeled with 125I by a modification of the Chloramine-T technique. The specific activity of the labeled hormone was 29 MBq/micrograms of AVP, as estimated by self- displacement analysis. The assay involves Sep-Pak C18 extraction of acidified (pH 4) plasma. Recovery of [3H]AVP added to plasma averaged 86.6 (SD 6.1)% (n = 14). Dilution curves for plasma showed linearity of response with concentration. The overall sensitivity was 0.3 ng/L when 2-mL plasma samples were extracted. The intra-assay CV was 7.8% at 4.8 ng/L (n = 12) and the interassay CV was 12.3% (n = 16) and 6.3% (n = 14) at 2.7 and 4.1 ng/L concentrations, respectively.


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M. Kluge, S. Riedl, B. Erhart-Hofmann, J. Hartmann, and F. Waldhauser
Improved Extraction Procedure and RIA for Determination of Arginine8-Vasopressin in Plasma: Role of Premeasurement Sample Treatment and Reference Values in Children
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Copyright © 1985 by the American Association for Clinical Chemistry.