I-125-desaminotyrosyl-β-alanin-secretin(I-125-DATA-secretin)を標識抗原とするsecretinのradioimmunoassay(RIA)の基礎的検討

概要

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A radioimmunoassay (RIA) for secretin using desaminotyrosyl-β-alanin-secretin (DATA-secretin) as a tracer and commercially available anti-secretin serum was developed. Separation of the bound and free forms was performed using the double antibody method. DATA-secretin was labelled with I-125 by use of the chloramine T method and radioactivities from the Bio Gel P-4 column of the labelled DATA-secretin were eluted in two peaks. I-125-DATA-secretin, which was reactive to the antiserum, was contained in the first peak, and the fractions on the down slope of I-125-DATA-secretin peak were the most suitable for the radioligand in the secretin RIA system. The specific activity of the purified I-125-DATA-secrestin ranged from 4.4 to 5.9 M Bq/μg. In the assay, the minimal detectable concentration of secretin was 5.4 pg/tube, and 54 pg/ml of plasma was possible to assay. Coefficient variations of the inter- and intrassay at 25 pg/tube were 13.7% and 7.8%, respectively. The mean recovery of the synthetic secretin added to the secretin free plasma was 96% at 500 pg/ml and 77% at 100 pg/ml. The incubation damage of the traccer was significantly lowered in pyrex tube as comparaed to plastic and polyethylen tubes. These results indicate that the RIA system using I-125-DATA-secretin as the tracer is sensitive and specific, and is useful for the physiological investigation of secretin.
公益社団法人日本放射線技術学会の論文
1985-01-01