Optimized LC-MS/MS Method for the Detection of ppCCK(21-44): A Surrogate to Monitor Human Cholecystokinin Secretion.
Optimized LC-MS/MS Method for the Detection of ppCCK(21-44): A Surrogate to Monitor Human Cholecystokinin Secretion.
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DOI：
10.1021/acs.jproteome.3c00272
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发表时间：
2023-09-01
影响因子：
4.4
通讯作者：
Kay, Richard G.
中科院分区：
文献类型：
--
作者：
Foreman, Rachel E.;Miedzybrodzka, Emily L.;Eiriksson, Finnur Freyr;Thorsteinsdottir, Margret;Bannon, Christopher;Wheller, Robert;Reimann, Frank;Gribble, Fiona M.;Kay, Richard G.
胆囊收缩素（CCK）是由十二指肠内分泌细胞在餐后分泌的，在低皮摩尔范围内循环。这种消化和食欲调节激素的检测目前依赖于免疫测定的使用，其中许多免疫测定在生理范围内的灵敏度不足，并且与以较高血浆浓度循环的胃泌素存在交叉反应性问题。作为现有技术的替代方案，开发了一种基于液相色谱和质谱的方法来测量细胞培养上清液中的CCK衍生肽。该方法最初应用于类器官研究，并且能够检测刺激后上清液中的CCK 8和N-末端肽片段（prepro）ppCCK（21-44）。使用统计建模软件进行提取优化，使得ppCCK（21-44）的定量LC-MS/MS方法能够在人血浆和分泌缓冲液中的低pM范围内检测该肽。分析了在过夜禁食后接受标准餐（Ensure）的健康个体的血浆样本;然而，该方法仅对检测ppCCK具有灵敏度（21-44）。在健康志愿者中采用人肠类器官的分泌研究和膳食研究证实，ppCCK（21-44）是用于测量CCK体外和体内释放的合适替代分析物。
The hormone cholecystokinin (CCK) is secreted postprandially from duodenal enteroendocrine cells and circulates in the low picomolar range. Detection of this digestion and appetite-regulating hormone currently relies on the use of immunoassays, many of which suffer from insufficient sensitivity in the physiological range and cross-reactivity problems with gastrin, which circulates at higher plasma concentrations. As an alternative to existing techniques, a liquid chromatography and mass spectrometry-based method was developed to measure CCK-derived peptides in cell culture supernatants. The method was initially applied to organoid studies and was capable of detecting both CCK8 and an N-terminal peptide fragment (prepro) ppCCK(21–44) in supernatants following stimulation. Extraction optimization was performed using statistical modeling software, enabling a quantitative LC-MS/MS method for ppCCK(21–44) capable of detecting this peptide in the low pM range in human plasma and secretion buffer solutions. Plasma samples from healthy individuals receiving a standardized meal (Ensure) after an overnight fast were analyzed; however, the method only had sensitivity to detect ppCCK(21–44). Secretion studies employing human intestinal organoids and meal studies in healthy volunteers confirmed that ppCCK(21–44) is a suitable surrogate analyte for measuring the release of CCK in vitro and in vivo.
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影响因子：
5.2
作者：
Rehfeld JF
通讯作者：
Rehfeld JF
影响因子：
--
作者：
Miedzybrodzka EL;Foreman RE;Galvin SG;Larraufie P;George AL;Goldspink DA;Reimann F;Gribble FM;Kay RG
通讯作者：
Kay RG
影响因子：
2.9
作者：
Rehfeld JF
通讯作者：
Rehfeld JF
影响因子：
7.7
作者：
Roberts, Geoffrey P.;Larraufie, Pierre;Gribble, Fiona M.
通讯作者：
Gribble, Fiona M.
影响因子：
7.4
作者：
Young, Scott A.;Julka, Samir;Yokoyama, Wallace H.
通讯作者：
Yokoyama, Wallace H.