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A sensitive enzyme immunoassay for atrial natriuretic polypeptide
H Watanabe1, M Wakimasu, K Kondo
1Chemistry Research Laboratories, Takeda Chemical Industries, Ltd., Osaka, Japan.
A sensitive sandwich enzyme immunoassay for mammalian alpha-atrial natriuretic polypeptide (alpha-ANP) has been developed using Fab' fragments of affinity purified rabbit anti-alpha-hANP[6-28] polyclonal antibodies (R3) conjugated to horseradish peroxidase. Immunoplates were coated with (F(ab')2) fragments of anti-alpha-hANP monoclonal antibodies (HA53). Without using a pre-concentration step the detection limit of alpha-hANP in plasma was 5 pg/ml. The assay was sufficient accuracy and reproducibility for general use of, i.e., the coefficients of within-assay and between-assay variation were 6.4-10.7% and 9.7-15.3%, respectively. Using the assay, the basal plasma alpha-hANP level of healthy persons was 15.3 +/- 8.3 pg/ml (mean +/- SD, n = 40).
A sensitive sandwich enzyme immunoassay for mammalian alpha-atrial natriuretic polypeptide (alpha-ANP) has been developed using Fab' fragments of affinity purified rabbit anti-alpha-hANP[6-28] polyclonal antibodies (R3) conjugated to horseradish peroxidase. Immunoplates were coated with (F(ab')2) fragments of anti-alpha-hANP monoclonal antibodies (HA53). Without using a pre-concentration step the detection limit of alpha-hANP in plasma was 5 pg/ml. The assay was sufficient accuracy and reproducibility for general use of, i.e., the coefficients of within-assay and between-assay variation were 6.4-10.7% and 9.7-15.3%, respectively. Using the assay, the basal plasma alpha-hANP level of healthy persons was 15.3 +/- 8.3 pg/ml (mean +/- SD, n = 40).