Harvey R F, Dowsett L, Hartog M, Read A E
Gut. 1974 Sep;15(9):690-9. doi: 10.1136/gut.15.9.690.
A sensitive and specific radioimmunoassay for cholecystokinin-pancreozymin (CCK-PZ) has been developed, using rabbit antisera to crude porcine hormone. Highly purified porcine CCK-PZ, labelled with (131)I, and repurified by column chromatography on Sephadex G15, was used as tracer. Separation of free from antibody-bound labelled CCK-PZ was carried out using charcoal, ion-exchange resin, or a double antibody procedure. Non-specific interference with the assay system by serum factors was abolished (as judged by in-vitro and in-vivo recovery studies) by boiling and diluting the serum samples before assay. Ninety-nine per cent pure porcine CCK-PZ (standard), commercial CCK-PZ preparations, caerulein, the C-terminal 8- and 12-amino acid fragments of the CCK-PZ molecule, and endogenous human CCK-PZ all cross reacted in the assay system and showed parallel inhibition curves. No significant cross reaction was found with gastrin, secretin, glucagon, or insulin. The sensitivity of the assay is approximately 5 pg per ml of test solution, which proved adequate for measuring physiological levels of CCK-PZ in peripheral blood in man.The mean immunoreactive CCK-PZ concentration in 50 fasting normal subjects was 60.4 pg per ml. The distribution of individual values was skewed, however, so that the median was much lower (30 pg per ml). Older subjects had higher fasting levels of CCK-PZ than were found in young adults.
已开发出一种用于胆囊收缩素-促胰酶素(CCK-PZ)的灵敏且特异的放射免疫测定法,该方法使用兔抗猪粗激素血清。用(131)I标记并经Sephadex G15柱色谱法再纯化的高度纯化的猪CCK-PZ用作示踪剂。使用活性炭、离子交换树脂或双抗体程序将游离的与抗体结合的标记CCK-PZ分离。通过在测定前将血清样品煮沸和稀释,消除了血清因子对测定系统的非特异性干扰(通过体外和体内回收率研究判断)。99%纯的猪CCK-PZ(标准品)、市售CCK-PZ制剂、蛙皮素、CCK-PZ分子的C末端8肽和12肽片段以及内源性人CCK-PZ在测定系统中均发生交叉反应,并呈现平行抑制曲线。与胃泌素、促胰液素、胰高血糖素或胰岛素未发现明显交叉反应。该测定法的灵敏度约为每毫升测试溶液5皮克,这证明足以测量人体外周血中CCK-PZ的生理水平。50名空腹正常受试者的平均免疫反应性CCK-PZ浓度为每毫升60.4皮克。然而,个体值的分布呈偏态,因此中位数要低得多(每毫升30皮克)。老年受试者的空腹CCK-PZ水平高于年轻人。