Radio-immunoassays for glucagon-like peptides 1 and 2 (GLP-1 and GLP-2).
Radio-immunoassays for glucagon-like peptides 1 and 2 (GLP-1 and GLP-2).
复制标题
胰高血糖素样肽 1 和 2（GLP-1 和 GLP-2）的放射免疫测定。
DOI：
10.1080/00365518709168885
复制
发表时间：
1987
影响因子：
2.1
通讯作者：
J. Holst
中科院分区：
文献类型：
--
作者：
C. Ørskov;J. Holst
基因测序研究表明，胰高血糖素前体含有两个额外的胰高血糖素样序列，即所谓的胰高血糖素样肽1和2（GLP-1和GLP-2）。我们开发了放射免疫分析对合成肽对应于这些序列。将肽与BSA碳二亚胺缀合后，在兔中产生抗血清。所选抗血清既不显示相互交叉反应性，也不与胰高血糖素家族的任何其他肽交叉反应。胰蛋白酶消化实验表明，这两种抗血清针对的C-末端的抗原肽。用氯胺T进行标记，并用等离子体包被的活性炭进行分离。GLP-1和GLP-2的检测限分别为7和25 pmol/l。血浆中两种肽的准确测量需要提取。乙醇浓度为75%（终浓度）时，回收率最佳。在10名正常受试者的空腹血浆中测得的GLP-1和GLP-2浓度分别为107 +/- 7 pmol/l和151 +/- 14 pmol/l。混合餐后，浓度缓慢上升2小时，达到145 +/- 13和225 +/- 15 pmol/l。
Gene-sequencing studies have shown that the glucagon precursor contains two additional glucagon-like sequences, the so-called glucagon-like peptides 1 and 2 (GLP-1 and GLP-2). We developed radio-immunoassays against synthetic peptides corresponding to these sequences. Antisera were raised in rabbits after carbodiimide conjugation of peptides to BSA. The selected antisera showed neither mutual cross-reactivity nor cross-reacted with any other peptide of the glucagon family. Trypsin digestion experiments showed that both antisera were directed against the C-terminus of the antigen peptides. Labelling was performed with chloramine T, and separation with plasma-coated charcoal. The detection limit was 7 and 25 pmol/l for GLP-1 and GLP-2. Accurate measurement of both peptides in plasma required extraction. Optimum recovery was obtained with ethanol at 75% (final concentration). The concentrations measured in fasting plasma from 10 normal subjects were 107 +/- 7 pmol/l and 151 +/- 14 pmol/l for GLP-1 and GLP-2, respectively. After a mixed meal the concentrations rose slowly for 2 h reaching 145 +/- 13 and 225 +/- 15 pmol/l.