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基于适配体共轭氧化石墨烯的表面辅助激光解吸电离质谱法用于阿尔茨海默病SH-SY5细胞模型中Aβ的选择性提取与检测

Aptamer-conjugated graphene oxide-based surface assisted laser desorption ionization mass spectrometry for selective extraction and detection of Aβ in an Alzheimer's disease SH-SY5 cell model.

作者信息

Song Gongshuai, Shui Ruofan, Wang Danli, Fang Ruosi, Yuan Tinglan, Li Ling, Feng Junli, Gao Feng, Shen Qing, Gong Jinyan, Zheng Fuping, Zhang Manman

机构信息

Zhejiang Provincial Key Lab for Biological and Chemical Processing Technologies of Farm Product, School of Biological and Chemical Engineering, Zhejiang University of Science and Technology, Hangzhou, China.

Zhejiang Province Joint Key Laboratory of Aquatic Products Processing, Collaborative Innovation Center of Seafood Deep Processing, Institute of Seafood, Zhejiang Gongshang University, Hangzhou, China.

出版信息

Front Aging Neurosci. 2022 Sep 20;14:993281. doi: 10.3389/fnagi.2022.993281. eCollection 2022.

Abstract

The generation and accumulation of amyloid-beta peptide (Aβ) in amyloid plaques are key characteristics of Alzheimer's disease (AD); thus, specific detection of Aβ is essential for the diagnosis and treatment of AD. Herein, an aptamer-conjugated graphene oxide (Apt-GO) sensor was synthesized by π-π and hydrophobic interactions using thiol poly (ethylene glycol) amine (SH-PEG-NH) as a spacer unit. Then, it was applied to selective capture of Aβ, and the resulting complex was directly analyzed by surface-assisted laser desorption ionization mass spectrometry (SALDI-MS). The results revealed that the Apt-GO could enhance the detection specificity and reduce non-specific adsorption. This method was validated to be sensitive in detecting Aβ at a low level in human serum (ca. 0.1 μM) within a linear range from 0.1 to 10 μM. The immobilizing amount of aptamer on the GO was calculated to be 36.1 nmol/mg (RSD = 11.5%). In conclusion, this Apt-GO-based SALDI-MS method was sensitive and efficient in selective extraction and detection of Aβ, which proved to be a good option for early AD diagnosis.

摘要

淀粉样β肽(Aβ)在淀粉样斑块中的产生和积累是阿尔茨海默病(AD)的关键特征;因此,Aβ的特异性检测对于AD的诊断和治疗至关重要。在此,使用硫醇聚(乙二醇)胺(SH-PEG-NH)作为间隔单元,通过π-π和疏水相互作用合成了一种适配体缀合的氧化石墨烯(Apt-GO)传感器。然后,将其应用于Aβ的选择性捕获,并通过表面辅助激光解吸电离质谱(SALDI-MS)直接分析所得复合物。结果表明,Apt-GO可以提高检测特异性并减少非特异性吸附。该方法经验证在检测人血清中低水平的Aβ(约0.1μM)时具有敏感性,线性范围为0.1至10μM。计算得出氧化石墨烯上适配体的固定量为36.1 nmol/mg(相对标准偏差=11.5%)。总之,这种基于Apt-GO的SALDI-MS方法在Aβ的选择性提取和检测中灵敏且高效,被证明是早期AD诊断的一个良好选择。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cd36/9530460/df6724b1a4bc/fnagi-14-993281-g001.jpg

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