Eye Institute and Department of Ophthalmology, Eye & ENT Hospital, Fudan University, Shanghai, 200031, China; NHC Key Laboratory of Myopia (Fudan University), Key Laboratory of Myopia, Chinese Academy of Medical Sciences, Shanghai Key Laboratory of Visual Impairment and Restoration, Shanghai, 200031, China; State Key Laboratory of Medical Neurobiology and MOE Frontiers Center for Brain Science, Institutes of Brain Science, Fudan University, Shanghai, 200032, China.
Eye Institute and Department of Ophthalmology, Eye & ENT Hospital, Fudan University, Shanghai, 200031, China; NHC Key Laboratory of Myopia (Fudan University), Key Laboratory of Myopia, Chinese Academy of Medical Sciences, Shanghai Key Laboratory of Visual Impairment and Restoration, Shanghai, 200031, China.
Biosens Bioelectron. 2022 Nov 1;215:114566. doi: 10.1016/j.bios.2022.114566. Epub 2022 Jul 14.
Although biosensors for signal monitoring have been extensively developed, their application in one-step high-throughput detection of low-abundance disease biomarkers remains challenging. This study presents a 3D aptasensor based on a biolayer interferometry (BLI) technique, followed by the sensitive and rapid detection of the specific biomarker brain-derived neurotrophic factor (BDNF) for early screening of glaucoma, an irreversible disease that causes blindness. The developed 3D aptasensor enabled one-step batch conversion of the low-abundance biomarker BDNF binding into optical interference signal, which was mainly attributed to the following factors: (1) A dimeric aptamer with extremely high targeting affinity was constructed as a biorecognition molecule, (2) highly sensitive 3D matrix sensors were integrated as signal transduction elements, and (3) the BLI Octet system with automated, high-throughput, and real-time online monitoring capabilities was used for reporting. The 3D aptasensor exhibited a broad detection window from 0.41 to 250 ng/mL BDNF, with a limit of detection of 0.2 ng/mL. Furthermore, detection of BDNF in glaucoma patient serum using the aptasensor showed good agreement with ELISA findings as well as the clinical diagnosis of the patient, demonstrating the feasibility of the system as a screening tool for glaucoma. This one-step high-throughput screening approach provides a valuable solution for the early diagnosis of glaucoma and may reduce the risk of blindness in visually impaired people.
尽管用于信号监测的生物传感器已经得到了广泛的发展,但它们在一步高通量检测低丰度疾病生物标志物方面的应用仍然具有挑战性。本研究提出了一种基于生物层干涉(BLI)技术的 3D 适体传感器,随后对特定生物标志物脑源性神经营养因子(BDNF)进行了灵敏快速的检测,用于早期筛查青光眼,这是一种不可逆的致盲疾病。所开发的 3D 适体传感器能够将低丰度生物标志物 BDNF 的结合一步批量转化为光学干涉信号,这主要归因于以下因素:(1)构建了具有极高靶向亲和力的二聚体适体作为生物识别分子,(2)集成了高灵敏度的 3D 基质传感器作为信号转导元件,以及(3)使用具有自动化、高通量和实时在线监测能力的 BLI Octet 系统进行报告。该 3D 适体传感器在 0.41 至 250ng/mL BDNF 范围内具有较宽的检测窗口,检测限为 0.2ng/mL。此外,使用适体传感器检测青光眼患者血清中的 BDNF 与 ELISA 结果以及患者的临床诊断具有良好的一致性,证明了该系统作为青光眼筛查工具的可行性。这种一步高通量筛选方法为青光眼的早期诊断提供了有价值的解决方案,可能降低视力受损人群失明的风险。