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关于阿皮林受体配体阿皮林测量方法的研究

On Methods for the Measurement of the Apelin Receptor Ligand Apelin.

机构信息

PKD Research Group, Laboratory of Pediatrics, Department of Development and Regeneration, KU Leuven, Leuven, Belgium.

Department of Nephrology and Arterial Hypertension, Universitair Ziekenhuis Brussel (UZ Brussel), Vrije Universiteit Brussel, Brussels, Belgium.

出版信息

Sci Rep. 2022 May 11;12(1):7763. doi: 10.1038/s41598-022-11835-z.

DOI:10.1038/s41598-022-11835-z
PMID:35546171
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9095593/
Abstract

Apelin exists in many isoforms, both in the circulation and in specific tissues. Apelin peptides have a short half-life but preservation before measurement is scarcely studied. Reproducible mass spectrometry methods to specifically measure a broad range of apelinergic peptide isoforms are currently lacking. A sample protocol to conserve apelinergic peptides in the preanalytical phase and a high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method to measure apelinergic isoforms was developed. Apelin was measured in plasma. For validation, human embryonic kidney (HEK) cells transfected with cDNA for preproapelin were used. Results were compared with a validated radioimmunoassay (RIA) method. Acidifying plasma to pH 2.5 improves post-sampling stability of apelin. HPLC-MS/MS was unable to detect apelin isoforms in plasma of healthy volunteers (n = 16) and chronic kidney disease patients (n = 4). RIA could detect apelin in concentrations between 71 and 263 fmol/l in 10 healthy volunteers. An optimized preanalytical protocol was developed. A sensitive and specific HPLC-MS/MS method failed to detect apelin in human plasma. Apelin-36 was detected in HEK cells transfected with cDNA for preproapelin. Currently, RIA with relatively selective antibodies is the best alternative for the measurement of apelin but novel sensitive and specific methods are needed.

摘要

Apelin 存在多种异构体，既存在于循环系统中，也存在于特定组织中。Apelin 肽半衰期短，但在测量前的保存情况很少被研究。目前缺乏可特异性测量广泛 Apelin 肽异构体的重现性质谱方法。本研究开发了一种在分析前阶段保存 Apelin 肽的样本方案和一种高效液相色谱-串联质谱 (HPLC-MS/MS) 方法来测量 Apelin 异构体。在血浆中测量 Apelin。为了验证，使用转染 cDNA 前体 Apelin 的人胚肾 (HEK) 细胞。结果与经过验证的放射免疫分析 (RIA) 方法进行比较。将血浆酸化至 pH 2.5 可改善 Apelin 采样后的稳定性。HPLC-MS/MS 无法检测健康志愿者（n = 16）和慢性肾脏病患者（n = 4）血浆中的 Apelin 异构体。RIA 可在 10 名健康志愿者中检测到 71 至 263 fmol/l 浓度的 Apelin。优化了分析前方案。灵敏且特异性的 HPLC-MS/MS 方法未能检测到人类血浆中的 Apelin。Apelin-36 在转染 cDNA 前体 Apelin 的 HEK 细胞中被检测到。目前，具有相对选择性抗体的 RIA 是测量 Apelin 的最佳选择，但需要新的灵敏且特异性方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dbf1/9095593/357b21dcb1ff/41598_2022_11835_Fig8_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dbf1/9095593/9b2066b5e883/41598_2022_11835_Fig1_HTML.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dbf1/9095593/011de8ae297b/41598_2022_11835_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dbf1/9095593/1448592c2122/41598_2022_11835_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dbf1/9095593/dfaccd67f5c1/41598_2022_11835_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dbf1/9095593/357b21dcb1ff/41598_2022_11835_Fig8_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dbf1/9095593/9b2066b5e883/41598_2022_11835_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dbf1/9095593/a05c4baf7079/41598_2022_11835_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dbf1/9095593/1473cc9a6d51/41598_2022_11835_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dbf1/9095593/00d4f3bfce58/41598_2022_11835_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dbf1/9095593/011de8ae297b/41598_2022_11835_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dbf1/9095593/1448592c2122/41598_2022_11835_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dbf1/9095593/dfaccd67f5c1/41598_2022_11835_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dbf1/9095593/357b21dcb1ff/41598_2022_11835_Fig8_HTML.jpg

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