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Novel and sensitive noncompetitive enzyme immunoassay hetero two site enzyme immunoassay for arginine vasopressin in plasma



Novel and sensitive noncompetitive enzyme immunoassay hetero two site enzyme immunoassay for arginine vasopressin in plasma



Analytical Letters 24(7): 1109-1124



A novel and sensitive noncompetitive enzyme immunoassay (hetero-two-site enzyme immunoassay) for arginine vasopressin in plasma is described. Plasma (0.3 ml) was diluted 1.3-fold with an appropriate buffer and filtered by centrifugation in a microconcentrator with polysaccharide membrane to eliminate plasma proteins. Arginine vasopressin in plasma filtrates was biotinylated and trapped onto anti-arginine vasopressin IgG-coated polystyrene balls. After washing the polystyrene balls to eliminate other biotinylated substances, the biotinylated arginine vasopressin was eluted from the polystyrene balls with HCl and was reacted with anti-arginine vasopressin Fab'-peroxidase conjugate. The complex formed was trapped onto streptavidin-coated polystyrene balls. Peroxidase activity bound to the polystyrene balls was assayed by fluorometry. The detection limit of arginine vasopressin was 11 fg (10 amol)/tube. This was 45-fold lower than that by competitive enzyme immunoassay during the same antiserum as used in this study and 9 to 400-fold lower than those previously reported by competitive radioimmunoassays. The assay range of arginine vasopressin in plasma was 0.14-140 ng/l using 100 .mu.l of plasma filtrates corresponding to 75 .mu.l of plasma. Plasma levels of arginine vasopressin in 8 healthy subjects aged 25-41 yr with ad libitum water intake and normal activity approximately 4 h after breakfast were 0.72 .+-. 0.22 (SD) ng/l (range, 0.42-1.04 ng/l).

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Accession: 007602583

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