Preanalytical impact on the accuracy of measurements of glucagon, GLP-1 and GIP in clinical trials.

IF 1.3 4区 医学 Q4 MEDICINE, RESEARCH & EXPERIMENTAL Scandinavian Journal of Clinical & Laboratory Investigation Pub Date : 2023-12-01 Epub Date: 2024-01-24 DOI:10.1080/00365513.2023.2294470
Christine Rasmussen, Michael M Richter, Nicole J Jensen, Niklas Heinz, Bolette Hartmann, Jens J Holst, Sasha A S Kjeldsen, Nicolai J Wewer Albrechtsen
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Abstract

Background: Plasma concentrations of glucagon, GLP-1 and GIP are reported in numerous clinical trials as outcome measures but preanalytical guidelines are lacking. We addressed the impact of commonly used blood containers in metabolic research on measurements of glucagon, GLP-1 and GIP in humans.

Methods: Seventeen overweight individuals were subjected to an overnight fast followed by an intravenous infusion of amino acids to stimulate hormonal secretion. Blood was sampled into five containers: EDTA-coated tubes supplemented with DMSO (control), a neprilysin inhibitor, aprotinin (a kallikrein inhibitor) or a DPP-4 inhibitor, and P800 tubes. Plasma was kept on ice before and after centrifugation and stored at -80 Celsius until batch analysis using validated sandwich ELISAs or radioimmunoassays (RIA).

Results: Measures of fasting plasma glucagon did not depend on sampling containers, whether measured by ELISA or RIA. Amino acid-induced hyperglucagonemia was numerically higher when blood was collected into P800 tubes or tubes with aprotinin. The use of p800 tubes resulted in higher concentrations of GLP-1 by RIA compared to control tubes but not for measurements with sandwich ELISA. Plasma concentrations of GIP measured by ELISA were higher in control tubes and negatively affected by P800 and the addition of aprotinin.

Conclusions: The choice of blood containers impacts on measurements of plasma concentrations of glucagon, GLP-1 and GIP, and based on this study, we recommend using EDTA-coated tubes without protease inhibitors or P800 tubes for measurements of glucagon, GLP-1 and GIP in clinical trials.

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分析前对临床试验中胰高血糖素、GLP-1 和 GIP 测量准确性的影响。
背景:许多临床试验都将胰高血糖素、GLP-1 和 GIP 的血浆浓度作为结果指标进行报告,但缺乏分析前指南。我们研究了代谢研究中常用的血液容器对人体胰高血糖素、GLP-1 和 GIP 测量的影响:方法:对 17 名超重者进行一夜禁食,然后静脉注射氨基酸以刺激荷尔蒙分泌。在五个容器中采血:EDTA涂层试管中分别加入二甲基亚砜(对照组)、肾蛋白酶抑制剂、阿普罗宁(凯利克林抑制剂)或DPP-4抑制剂,以及P800试管。血浆在离心前后均置于冰上,并储存在零下 80 摄氏度的环境中,直到使用经过验证的夹心酶联免疫吸附试验或放射免疫分析法(RIA)进行批量分析:结果:无论是采用酶联免疫吸附法还是放射免疫分析法,空腹血浆胰高血糖素的测量值都与采样容器无关。用 P800 管或含阿普丁的管采血时,氨基酸诱发的高胰高血糖素血症在数量上更高。与对照试管相比,使用p800试管通过RIA测定的GLP-1浓度更高,但使用夹心ELISA测定的GLP-1浓度则不高。用酶联免疫吸附法测定的血浆中 GIP 浓度在对照试管中更高,而 P800 和添加阿普罗宁会对其产生负面影响:血液容器的选择会影响胰高血糖素、GLP-1 和 GIP 的血浆浓度测量,根据这项研究,我们建议在临床试验中使用不含蛋白酶抑制剂的 EDTA 涂层试管或 P800 试管来测量胰高血糖素、GLP-1 和 GIP。
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来源期刊
CiteScore
3.50
自引率
4.80%
发文量
85
审稿时长
4-8 weeks
期刊介绍: The Scandinavian Journal of Clinical and Laboratory Investigation is an international scientific journal covering clinically oriented biochemical and physiological research. Since the launch of the journal in 1949, it has been a forum for international laboratory medicine, closely related to, and edited by, The Scandinavian Society for Clinical Chemistry. The journal contains peer-reviewed articles, editorials, invited reviews, and short technical notes, as well as several supplements each year. Supplements consist of monographs, and symposium and congress reports covering subjects within clinical chemistry and clinical physiology.
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