Synthesis and characterization of miniaturized aptamer-based monolithic sorbent for selective extraction of β-amyloid peptides from cerebrospinal fluid.

IF 3.8 2区 化学 Q1 BIOCHEMICAL RESEARCH METHODS
Israel Donizeti de Souza, Caroline Fernandes Grecco, Maria Eugênia Costa Queiroz, Valerie Pichon, Audrey Combès
{"title":"Synthesis and characterization of miniaturized aptamer-based monolithic sorbent for selective extraction of β-amyloid peptides from cerebrospinal fluid.","authors":"Israel Donizeti de Souza, Caroline Fernandes Grecco, Maria Eugênia Costa Queiroz, Valerie Pichon, Audrey Combès","doi":"10.1007/s00216-025-06085-7","DOIUrl":null,"url":null,"abstract":"<p><p>The ratio between beta-amyloid (Aβ) peptides 40 and 42 is recognized as a biomarker for Alzheimer's disease, playing a significant role in early diagnosis and disease progression monitoring. Aβ peptides are present at trace levels in cerebrospinal fluid, therefore, developing a new selective extraction procedure is essential for isolating targeted biomarkers from the matrix interferents, ensuring accurate identification and quantification. In this study, a hybrid organic-silica monolith was synthesized in a 530 µm inner diameter-capillary and used for the covalent grafting of beta amyloid peptide aptamers. The resulting miniaturized oligosorbent (mOS) was applied to selectively extract Aβ peptides 40 and 42 from artificial cerebrospinal fluid (CSF) samples. The immobilization procedure achieved grafting yields higher than 90% leading to a dense coverage of Aβ aptamers (655 + 15 pmol.µL<sup>-1</sup> of oligosorbent, n = 3, RSD = 2.3), and in a capacity exceeding 50 pmol.µL<sup>-1</sup> of mOS. After optimization in pure media, an extraction recovery of 74% and 31% for Aβ40 and Aβ42 peptides, respectively was reached on this mOS. The developed method including extraction on mOS and LC-MS analysis achieved LLOQ values of 0.1 ng.mL<sup>-1</sup>, precision and accuracy with CV and RSD values ranging from 1.0% to 12.9% and -4.7% to 11.1%, respectively. This method was successfully applied to selectively extract Aβ peptides from artificial CSF samples, effectively isolating the two Aβ targeted peptides from this complex biological fluid and enhancing trace-level analysis reliability.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8000,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Analytical and Bioanalytical Chemistry","FirstCategoryId":"92","ListUrlMain":"https://doi.org/10.1007/s00216-025-06085-7","RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOCHEMICAL RESEARCH METHODS","Score":null,"Total":0}
引用次数: 0

Abstract

The ratio between beta-amyloid (Aβ) peptides 40 and 42 is recognized as a biomarker for Alzheimer's disease, playing a significant role in early diagnosis and disease progression monitoring. Aβ peptides are present at trace levels in cerebrospinal fluid, therefore, developing a new selective extraction procedure is essential for isolating targeted biomarkers from the matrix interferents, ensuring accurate identification and quantification. In this study, a hybrid organic-silica monolith was synthesized in a 530 µm inner diameter-capillary and used for the covalent grafting of beta amyloid peptide aptamers. The resulting miniaturized oligosorbent (mOS) was applied to selectively extract Aβ peptides 40 and 42 from artificial cerebrospinal fluid (CSF) samples. The immobilization procedure achieved grafting yields higher than 90% leading to a dense coverage of Aβ aptamers (655 + 15 pmol.µL-1 of oligosorbent, n = 3, RSD = 2.3), and in a capacity exceeding 50 pmol.µL-1 of mOS. After optimization in pure media, an extraction recovery of 74% and 31% for Aβ40 and Aβ42 peptides, respectively was reached on this mOS. The developed method including extraction on mOS and LC-MS analysis achieved LLOQ values of 0.1 ng.mL-1, precision and accuracy with CV and RSD values ranging from 1.0% to 12.9% and -4.7% to 11.1%, respectively. This method was successfully applied to selectively extract Aβ peptides from artificial CSF samples, effectively isolating the two Aβ targeted peptides from this complex biological fluid and enhancing trace-level analysis reliability.

用于脑脊液β-淀粉样肽选择性提取的小型适配体整体吸附剂的合成与表征。
β -淀粉样蛋白(a β)肽40和42的比值被认为是阿尔茨海默病的生物标志物,在早期诊断和疾病进展监测中发挥重要作用。a β肽在脑脊液中以痕量水平存在,因此,开发一种新的选择性提取方法对于从基质干扰物中分离靶向生物标志物,确保准确鉴定和定量至关重要。本研究在内径530µm的毛细管中合成了有机硅杂化单体,并将其用于β -淀粉样肽适体的共价接枝。应用微型寡吸附剂(mOS)从人工脑脊液(CSF)样品中选择性地提取Aβ肽40和42。固定化过程实现了高于90%的接枝率,导致a β适配体的密集覆盖(655 + 15 pmol)。µL-1的低聚吸附剂,n = 3, RSD = 2.3),容量超过50 pmol。µL-1的mOS。经纯培养基优化后,Aβ40和Aβ42肽的提取率分别为74%和31%。所开发的方法包括mOS提取和LC-MS分析,LLOQ值为0.1 ng。精密度和准确度的CV和RSD值分别为1.0% ~ 12.9%和-4.7% ~ 11.1%。该方法成功地从人工脑脊液样品中选择性地提取了Aβ肽,有效地从复杂的生物液中分离出了两种Aβ靶向肽,提高了痕量水平分析的可靠性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
CiteScore
8.00
自引率
4.70%
发文量
638
审稿时长
2.1 months
期刊介绍: Analytical and Bioanalytical Chemistry’s mission is the rapid publication of excellent and high-impact research articles on fundamental and applied topics of analytical and bioanalytical measurement science. Its scope is broad, and ranges from novel measurement platforms and their characterization to multidisciplinary approaches that effectively address important scientific problems. The Editors encourage submissions presenting innovative analytical research in concept, instrumentation, methods, and/or applications, including: mass spectrometry, spectroscopy, and electroanalysis; advanced separations; analytical strategies in “-omics” and imaging, bioanalysis, and sampling; miniaturized devices, medical diagnostics, sensors; analytical characterization of nano- and biomaterials; chemometrics and advanced data analysis.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:604180095
Book学术官方微信