Labeling tumor-associated extracellular vesicles with antibody-DNA conjugates for quantitative analysis.

IF 3.9 3区 生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY
Frontiers in Molecular Biosciences Pub Date : 2025-01-22 eCollection Date: 2025-01-01 DOI:10.3389/fmolb.2025.1531108
Xiao Du, Hongxiu Li, Shiyi Shen, Chao Tian, Xiaohuan Cao, Xingang Xu, Nan Xu, Shuling Wang, Qingchang Tian
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引用次数: 0

Abstract

Introduction: Extracellular vesicles (EVs) shed from tumor cells into peripheral circulation or other body fluids are promising biomarkers for cancer diagnosis with enormously long circulation. Consequently, precise methods for differentiating normal and tumor-associated EVs (TAEs) are required.

Methods: This study used quantifiable antibody-DNA conjugate-assisted quantitative methods combined with proximity ligation technology to detect TAEs. The antibody-DNA conjugate contained one antibody associated with three oligonucleotides for signal amplification. The antibody in the conjugate can recognize the surface tumor antigens of TAEs. Simultaneously, DNA in the conjugate is attached to the surfaces of TAEs and holds the signal amplification post, converting protein identities to DNA amplification for protein detection, even at the molecular level.

Results: These findings revealed that TAEs can be quantitatively detected using DNA-mediated quantitative polymerase chain reaction (qPCR). Antibody-DNA conjugates were used to recognize the epithelial cell adhesion molecule (EpCAM) antigen on the TAE surface and quantify the antigen using qPCR for cancer analysis.

Discussion: This method proposed a new quantitative detection approach for TAEs, which aim to identify specific EV-associated markers for diagnostic or therapeutic, this method could inspire a new idea for tumor diagnosis and detection of other diseases.

用抗体- dna偶联物标记肿瘤相关的细胞外囊泡进行定量分析。
导读:细胞外囊泡(Extracellular vesicles, EVs)从肿瘤细胞中脱落到外周循环或其他体液中,是一种很有希望用于肿瘤诊断的生物标志物。因此,需要精确的方法来区分正常和肿瘤相关的ev (TAEs)。方法:采用定量抗体- dna偶联辅助定量方法结合近端结扎技术检测TAEs。该抗体- dna偶联物含有一个与三个寡核苷酸相关的抗体,用于信号扩增。结合物中的抗体能识别TAEs的表面肿瘤抗原。同时,缀合物中的DNA附着在TAEs表面并保持信号扩增柱,将蛋白质身份转换为DNA扩增以用于蛋白质检测,甚至在分子水平上也是如此。结果:利用dna介导的定量聚合酶链反应(qPCR)可以对TAEs进行定量检测。抗体- dna偶联物用于识别TAE表面的上皮细胞粘附分子(EpCAM)抗原,并使用qPCR定量抗原用于癌症分析。讨论:该方法提出了一种新的TAEs定量检测方法,旨在鉴定特异性的ev相关标志物,用于诊断或治疗,该方法可为肿瘤诊断和其他疾病的检测提供新的思路。
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来源期刊
Frontiers in Molecular Biosciences
Frontiers in Molecular Biosciences Biochemistry, Genetics and Molecular Biology-Biochemistry
CiteScore
7.20
自引率
4.00%
发文量
1361
审稿时长
14 weeks
期刊介绍: Much of contemporary investigation in the life sciences is devoted to the molecular-scale understanding of the relationships between genes and the environment — in particular, dynamic alterations in the levels, modifications, and interactions of cellular effectors, including proteins. Frontiers in Molecular Biosciences offers an international publication platform for basic as well as applied research; we encourage contributions spanning both established and emerging areas of biology. To this end, the journal draws from empirical disciplines such as structural biology, enzymology, biochemistry, and biophysics, capitalizing as well on the technological advancements that have enabled metabolomics and proteomics measurements in massively parallel throughput, and the development of robust and innovative computational biology strategies. We also recognize influences from medicine and technology, welcoming studies in molecular genetics, molecular diagnostics and therapeutics, and nanotechnology. Our ultimate objective is the comprehensive illustration of the molecular mechanisms regulating proteins, nucleic acids, carbohydrates, lipids, and small metabolites in organisms across all branches of life. In addition to interesting new findings, techniques, and applications, Frontiers in Molecular Biosciences will consider new testable hypotheses to inspire different perspectives and stimulate scientific dialogue. The integration of in silico, in vitro, and in vivo approaches will benefit endeavors across all domains of the life sciences.
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