Japanese
Title二抗体法RadioimmunoassayにおけるBound percentに及ぼす諸因子について
Subtitle原著
Authors吉村学*, 越智幸男*, 八谷孝*, 塩見勝彦*, 宮崎忠芳*, 岡本邦雄**
Authors(kana)
Organization*京都府立医科大学 第二内科, **同位元素研究室
Journal核医学
Volume10
Number5
Page451-456
Year/Month1973/10
Article原著
Publisher日本核医学会
Abstract「はじめに」Radioimmunoassay(RIA)はすでに一般化して各種のassay kitが各社より発売されている. 各assay kitには測定操作法が指定され, また良好な標準曲線も例示されている. しかし実際にassay kitを使用して標準曲線を作つてみるとそのbound percent(B%)は低い場合もあり, また必ずしも信頼できる値が得られないこともあるため, 各測定者はおのおの工夫を加えねばならないこともある. このたびわれわれは市販されているダイナボットRI研究所の成長ホルモン(HGH)測定用キットと第一ラジオアイソトープ研究所の甲状腺刺激ホルモン(HTSH)測定用キットを用いて被検血清によるB%低下因子の問題と二抗体法における第一抗体および第二抗体の量の問題につき検討したので報告する.
Practice臨床医学：一般
Keywords
English
TitleNon-specific inhibition of serum for bound percent in double antibody radioimmunoassay
SubtitleOriginal
AuthorsManabu YOSHIMURA, Yukio OCHI, Takashi HACHIYA, Katsuhiko SHIOMI, Tadayoshi MIYAZAKI, Kunio OKAMOTO
Authors(kana)
Organization2nd Department of Internal Medicine, Kyoto Prefectural University of Medicine, and Radioisotope Laboratory
JournalThe Japanese Journal of nuclear medicine
Volume10
Number5
Page451-456
Year/Month1973/10
ArticleOriginal article
PublisherTHE JAPANESE SOCIETY OF NUCLEAR MEDICINE
Abstract[Summary] By the development of radioimmunoassay (RIA) procedure, the assay procedure is established for clinical uses, and commercial Kitts for HGH and HTSH RIA (double antibody method) have been supplied from Dainabot RI laboratory and Daiichi RI laboratory. Even following the instructions manual precisely, it is very hard to get the exact values of each hormones, because the standard curve of each hormones were constructed using BSA (bovine serum albumin) buffer solution instead of diluent consisting of hormone free serum. In the present study, the effect of various human serum on the standard curve of HGH and HTSH RIA was analysed using pooled human serum as normal human serum (NHS), serum from the patient of panhypopituitarism (PPS) and the serum removed its hormone by dextran-coated charcoal (DCC) solution (20% charcoal and 2% dextran in saline). In figuring the standard curve of HGH and HTSH RIA, the bound percent (B%) of the standard curve using 0.5% BSA buffer solution (for HGH) or 1% BSA buffer solution (for HTSH) was higher than the B% of assay system containing with PPS, PPS treated with DCC, NHS and NHS treated with DCC. The above finding indicated that some factor to inhibit B% nonspecifically existed in serum, that the inhibiting factor of B% could not removed by DCC, and that the hormone free serum (HFS) is necessary for figuring the standard curve of RIA. In analysing the factor to inhibit B% in serum, the standard curves of HTSH with NHS or with NHS removed the complement by heat treatment with 56℃ for 30 min was figured identically and it was found that complement did not act on inhibiting factor for B%. Regarding the amount of first antibody (antiserum for HGH or HTSH) and of second antibody (anti-rabbit γ-globulin goat serum), different amounts of antibody, such as first antibody rich or second antibody rich, were given the three different assay system of serum free system, serum 0.1 ml and serum 0.3 ml using NHS, PPS and NHS treated with DCC. In this study it was found that B% in system containing serum was lower than that without serum. The B% in serum 0.3 ml was much lower than that in serum 0.1 ml. By appling larger amount of 2 nd antibody than the amount of instruction manual, it is possible to prohibit the decrease of B% inducing by serum 0.1 ml. However it is impossible to prohibit the decrease of B% in serum 0.3ml. On the contrary, the amount of 1 st antibody is also important to prohibit the decrease of B% in serum and both antibodies in optimal amounts are necessary to get the better result. From these results, it is very important to use hormone free serum as diluent for double antibody RIA.
PracticeClinical medicine
Keywords

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