| Japanese |
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| Title | GlucagonのRadioimmunoassayにおける基礎的検討 |
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| Subtitle | 原著 |
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| Authors | 村田和平*, 杉山陽一*,** |
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| Organization | *三重大学医学部産科婦人科学教室, **主任 教授 |
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| Journal | 核医学 |
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| Volume | 15 |
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| Number | 7 |
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| Page | 959-968 |
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| Year/Month | 1978/10 |
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| Article | 原著 |
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| Publisher | 日本核医学会 |
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| Abstract | 「要旨」: グルカゴンの免疫学的測定法において基礎的検討を行い, 新しい測定系を確立した. 結合型, 遊離型グルカゴンの分離においてPEG法はtalc法, 2抗体法, dextran coated charcoal法の3方法に比し操作が容易であり, しかも精度が優れた. 結合率はPEGの最終濃度が10.0%〜12.5% (w/w) で最高であった. まず検体と抗体を4℃で48時間前に温浴させ, その後標識グルカゴンを添加しさらに70時間行う2段法の導入により標準曲線の低濃度域における勾配が高まり, 感度および精度が向上した. 以上の成績から確立した2段法およびPEG法を用いた測定系の最小検出感度は16pg/ml, 再現性はIntraassayの変動係数1.9%〜5.4%, Interassayでは6.3%〜11.3%, 平均回収率は96.7%, 稀釈曲線では2〜8倍稀釈で直線性を示した. この測定系は感度, 信頼性が高く, 臨床上の診断に有用と考えられた. |
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| Practice | 臨床医学：一般 |
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| Keywords | Radioimmunoassay, Glucagon, Bound-free separation, Preinculation. |
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| English |
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| Title | Fundamental Research on Glucagon Radioimmunoassay |
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| Subtitle | Original Articles |
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| Authors | Kazuhira MURATA, Youichi Sugiyama1) |
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| Authors(kana) | |
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| Organization | Department of Obstetrics and Gynecology, Mie University, School of Medicine, 1)Director:Professor |
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| Journal | The Japanese Journal of nuclear medicine |
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| Volume | 15 |
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| Number | 7 |
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| Page | 959-968 |
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| Year/Month | 1978/10 |
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| Article | Original article |
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| Publisher | THE JAPANESE SOCIETY OF NUCLEAR MEDICINE |
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| Abstract | [Summary] Various factors and techniques concerning the glucagon radioimmunoassay were extensively studied and a new assay system was devised. The antigen-antibody reaction reached a plateau of maximal binding after 48 hours of incubation at 4℃. The uses of PEG, talc, double antibody and dextran coated charcoal to separate free and bound glucagon were compared. Although each method was possible to measure the plasma glucagon, PEG method was most convenient and satisfactory in precision in both standard and plasma samples. Time interval between addition of PEG and centrifugation showed no detectable effect. The net percent of immunoprecipitation was maximum at the PEG concentration of 10.0% - 12.5%. By means of preincubation of samples with antiserum for 48 hours before addition of 125I-glucagon, the slope of standard curve was sharpend at lower levels of glucagon. As the result, the sensitivity and precision were improved. The established radioimmunoassay procedure is as follows. The mixture of standard solution or unknown sample, 0.1 ml ; antiserum diduted (1/2,000) in phosphate buffered saline or buffer alone, 0.05 ml ; aprotinine, 0.05 ml ; and glycine buffer, 0.2 ml was preincubated for 48 hours. Following 0.1 ml of 125I-glucagon (125 pg/ml) was added and the mixture was incubated for 70 hours. It was necessary to adjust protein content to the same level in all tubes before precipitation of the glucagon-antibody complex. Polyethylene glycol #6,000 (final dilution 12.5%) was dispensed into all tubes. Then the tubes were centrifuged and the supernatant was aspirated. Sensitivity of this assay was 16 pg/ml over the range of 0 - 31.3pg/ml, and 25pg/ml over the range of 250pg/ml - 500pg/ml. Precision for intraassay was 1.9% - 5.4% (C.V.), and for interassay 6.3% - 11.3%. Recovery of exogenous glucagon added to plasma was satisfactory. Measurement of glucagon was possible in such low level as 1/8 of basal levels. This assay system, utilizing 2 stage incubation technique and PEG for separation of bound from free fraction, is very sensitive and reliable and considered very useful for clinical study. |
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| Practice | Clinical medicine |
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| Keywords | Radioimmunoassay, Glucagon, Bound-free separation, Preinculation. |
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