<br>脑源性神经营养因子 (BDNF) 靶标的高亲和力 DNA 适体的分离和鉴定,ACS Chemical Neuroscience

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脑源性神经营养因子 (BDNF) 靶标的高亲和力 DNA 适体的分离和鉴定
ACS Chemical Neuroscience ( IF 4.1 ) Pub Date : 2023-12-27 , DOI: 10.1021/acschemneuro.3c00661
Md Anisuzzaman Chowdhury ₁ , Jessica M Collins ₁ , David A Gell ₂ , Sharn Perry ₁ , Michael C Breadmore ₃ , Sarah Shigdar ₄ , Anna E King ₁

Affiliation

适体是用于各种靶标的分子识别的功能性寡核苷酸配体。适体的天然特性使其成为诊断、治疗和生物传感领域抗体的绝佳替代品。 DNA适体主要是具有与靶标明确结合的单链寡核苷酸（ssDNA）。然而，适体在脑健康和神经退行性疾病领域的应用迄今为止还受到限制。在此，通过体外筛选获得了针对脑源性神经营养因子（BDNF）蛋白的DNA适体。 BDNF 是大脑健康和神经退行性疾病的潜在生物标志物，在神经元的突触可塑性和存活中发挥作用。我们从 50 个核苷酸的文库中鉴定出 8 个对 BDNF 具有结合亲和力的适体。在这些适体中，NV_B12 显示出检测 BDNF 的最高灵敏度和选择性。在适配体连接的固定化吸附剂测定 (ALISA) 中，NV_B12 适配体以剂量依赖性方式与 BDNF 蛋白强烈结合。 NV_B12 的解离常数 ( K _d ) 为 0.5 nM (95% CI: 0.4–0.6 nM)。这些发现表明 BDNF 特异性适体可以用作 BDNF 诊断和检测测定中抗体的替代品。

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Isolation and Identification of the High-Affinity DNA Aptamer Target to the Brain-Derived Neurotrophic Factor (BDNF)

Aptamers are functional oligonucleotide ligands used for the molecular recognition of various targets. The natural characteristics of aptamers make them an excellent alternative to antibodies in diagnostics, therapeutics, and biosensing. DNA aptamers are mainly single-stranded oligonucleotides (ssDNA) that possess a definite binding to targets. However, the application of aptamers to the fields of brain health and neurodegenerative diseases has been limited to date. Herein, a DNA aptamer against the brain-derived neurotrophic factor (BDNF) protein was obtained by in vitro selection. BDNF is a potential biomarker of brain health and neurodegenerative diseases and has functions in the synaptic plasticity and survival of neurons. We identified eight aptamers that have binding affinity for BDNF from a 50-nucleotide library. Among these aptamers, NV_B12 showed the highest sensitivity and selectivity for detecting BDNF. In an aptamer-linked immobilized sorbent assay (ALISA), the NV_B12 aptamer strongly bound to BDNF protein, in a dose-dependent manner. The dissociation constant (K_d) for NV_B12 was 0.5 nM (95% CI: 0.4–0.6 nM). These findings suggest that BDNF-specific aptamers could be used as an alternative to antibodies in diagnostic and detection assays for BDNF.

更新日期：2023-12-27

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