Kominami G, Mori S, Sagawa K, Igano K, Inouye K, Kono M
Shionogi Research Laboratories, Shionogi & Co., Ltd., Osaka, Japan.
J Immunoassay. 1988;9(3-4):229-43. doi: 10.1080/01971528808053214.
A highly specific and sensitive competitive radioimmunoassay was developed for caerulein (CLN), an analogue of cholecystokinin-8 (CCK-8), in plasma and brain. Antiserum was produced in rabbit by immunization with N delta-[CLN-(1-6)]-ornithine amide conjugated with bovine serum albumin by the glutaraldehyde method. N alpha-[CLN-(1-6)]-lysine amide was labelled with 125I-Bolton & Hunter reagent and used as a labelled antigen after purification by high-performance liquid chromatography. This assay was highly specific for CLN, and cross reactivities for other related peptides, CCK-4, CCK-8, gastrin-I, and gastrin-(14-17), were not observed (less than 0.01%). The limits of determination in biological specimens after CLN administration were 11 pg/ml in human plasma and rat plasma and 80 pg/g in rat brain. This study showed that the slight structure difference between hapten and 125I-labelled antigen is important to the assay performance.
我们开发了一种高特异性和高灵敏度的竞争性放射免疫分析法，用于检测血浆和脑组织中的蛙皮素（CLN），它是胆囊收缩素-8（CCK-8）的类似物。通过戊二醛法用与牛血清白蛋白偶联的Nδ-[CLN-(1-6)]-鸟氨酸酰胺免疫家兔制备抗血清。用125I-博尔顿-亨特试剂标记Nα-[CLN-(1-6)]-赖氨酸酰胺，并在通过高效液相色谱纯化后用作标记抗原。该分析法对CLN具有高度特异性，未观察到对其他相关肽（CCK-4、CCK-8、胃泌素-I和胃泌素-(14-17)）的交叉反应（小于0.01%）。给予CLN后，生物样本中的检测限在人血浆和大鼠血浆中为11 pg/ml，在大鼠脑组织中为80 pg/g。这项研究表明，半抗原与125I标记抗原之间的微小结构差异对分析性能很重要。
