Ito K, Goto T, Tsuji A, Maeda M
School of Pharmaceutical Sciences, Showa University, Tokyo, Japan.
J Pharm Biomed Anal. 1997 Jun;15(9-10):1489-95. doi: 10.1016/s0731-7085(96)01985-1.
Pituitary adenylate cyclase activating polypeptide (PACAP) is a novel peptide hormone and has a variety of biological action. In studies of the physiological behaviour of endogenous PACAP, the determination of PACAP levels in biological materials require a highly sensitive and specific method. Therefore, we developed a sensitive time-resolved fluoroimmunoassay (TR-FIA) for PACAP27 which is the biologically important fragment of PACAP. Accordingly, we developed TR-FIA using a biotinylated PACAP27 (b-PACAP27) as a tracer and europium (III) chelate labeled streptavidin-biotinylated bovine serum albumin complex as a detection of biotin on solid phase. A measurable range of PACAP27 was 7.8-1000 pg m1-1 by the proposed TR-FIA. For measurement of biological samples, the samples were purified to eliminate substances which interfered with the TR-FIA. The mean recovery of PACAP27 using commercially reversed phase column was 74.7% (n = 12). The various tissues, extracts and plasma concentrations of rat could be measured by the proposed TR-FIA.
垂体腺苷酸环化酶激活多肽（PACAP）是一种新型肽类激素，具有多种生物学作用。在内源性PACAP生理行为的研究中，生物材料中PACAP水平的测定需要一种高度灵敏且特异的方法。因此，我们开发了一种针对PACAP27的灵敏时间分辨荧光免疫分析方法（TR-FIA），PACAP27是PACAP具有生物学重要性的片段。相应地，我们以生物素化的PACAP27（b-PACAP27）作为示踪剂，以铕（III）螯合物标记的链霉亲和素-生物素化牛血清白蛋白复合物作为固相生物素的检测物，开发了TR-FIA。通过所提出的TR-FIA方法，PACAP27的可测量范围为7.8 - 1000 pg m1-1。对于生物样品的测量，将样品进行纯化以去除干扰TR-FIA的物质。使用商业反相柱时，PACAP27的平均回收率为74.7%（n = 12）。所提出的TR-FIA方法能够测量大鼠的各种组织、提取物和血浆浓度。
