シロサケ成長ホルモンの競合法による酵素免疫測定法〔英文〕

概要

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An enzyme immunoassay (EIA), based on a competitive assay system, for the measurement of growth hormone (GH) in the pituitary of salmonid fishes and for the hormone released into pituitary incubation medium was developed usíng a rabbit antiserum against natural chum salmon growth hormone (nsGH I) isolated from pituitary glands. Recombinant salmon GH (rsGH I), was coupled to horseradish peroxidase (HRP). Antibody-bound HRP-rsGH I was separated from free HRP-rsGH I by a double antibody precipitation method. The enzyme activity in the precipitate was detected by a colorimetric method, in which o-phenylenediamine and hydrogen peroxide were used as substrates. nsGH I and rsGH I showed different affinity to the antibody. The displacement curves for extracts from chum salmon and rainbow trout pituitaríes, and incubation medium of the pars distalis of the trout were parallel to the GH standard, indicating that the present GH-EIA could estimate GH concentrations in those solutions. An alkaline solution was more effective than saline to extract GH from the pituitary gland. Among the chum salmon adenohy-phophyseal hormones examined, only gonadotropin I showed slight cross-reactivity (less than 0.1%). Neurohypophyseal hormones such as arginine vasotocin and isotocin showed slight (1%) and parallel cross-reactivities to the standard hormone.
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