A nanosized magnetic particle system for fast and efficient neuronal extracellular vesicle enrichment from plasma

摘要/摘要 胞外囊泡(EV)是纳米囊泡，它包含表面标记，并具有细胞和细胞状态特异性 其来源的组织(和细胞)的货物，反映组织或细胞的生理状态。最重要的是， 神经元电动汽车可以通过血脑屏障进入血液循环，并提供了一种新的途径 直接接触到大脑的致病过程。新出现的临床研究表明，神经元EVS 血液中携带致病生物标志物，可以检测和预测神经疾病，如阿尔茨海默氏症 疾病，帕金森病，肌萎缩侧索硬化症，创伤性脑损伤等。因此， 治疗神经系统疾病的脑组织成分。释放神经元电动汽车的巨大潜力 神经疾病诊断应用，快速高效地从血浆中浓缩神经元EVS 是一个先决条件。然而，有效地从血浆中分离神经元EV是一项重大挑战， 而且还没有商业上可用的技术/试剂盒来从血液中分离神经元EV 血浆。 在此，我们建议开发一种神经元EVS纳米探针(Neuron-NPs)技术，用于高效分离 神经元EV直接从血浆样本中作为研究用试剂盒。我们将开发和优化这些神经元- NPS，用于在缓冲系统中快速有效地磁隔离神经元EVS(目标1)。目标2将执行 血浆样品中神经元核蛋白技术的分析验证。这些目标的成功实现将产生 先进的专利技术平台，用于各种基础和临床研究中的神经元EVS分离 神经系统疾病。第二阶段的活动将集中在扩大神经元电动汽车的制造规模。 纳米探针试剂盒和开发与神经元集成的高通量自动样品处理系统- NPS套件。

SUMMARY/Abstract Extracellular vesicles (EVs) are nanovesicles, which contain surface markers, and cell- and cell-state-specific cargo of their tissue (and cell) of origin, reflecting the tissue or cell’s physiological state. Most importantly, neuronal EVs can pass the blood-brain barrier (BBB) into blood circulation and provide a new approach of gaining direct access to brain pathogenic processes. Emerging clinical studies show that neuronal EVs in blood carry pathogenic biomarkers that can detect and predict neurological diseases such as, Alzheimer’s disease, Parkinson disease, amyotrophic lateral sclerosis, and traumatic brain injury, etc. Therefore, enrichment of neuronal EVs from peripheral blood provides a minimally invasive way to sample components of brain tissue for neurological diseases. To unleash neuronal EVs’ great potential in neurological diseases diagnosis applications, rapid and efficient enrichment of neuronal EVs from blood plasma is a prerequisite. However, there is major challenge to efficiently isolate neuronal EVs from blood plasma, and there are no commercially available technologies/kits for the isolation of neuronal EVs from blood plasma. Herein, we propose to develop a neuronal EVs nanoprobes (neuron-NPs) technology for high-yield isolation of neuronal EV directly from plasma sample as a research use only kit. We will develop and optimize these neuron- NPs for rapid and efficient magnetic isolation of neuronal EVs in a buffer system (Aim 1). The Aim 2 will perform analytical validation of neuron-NP technology in plasma samples. Successful completion of these Aims will yield an advanced proprietary technology platform for neuronal EVs isolations in basic and clinical research for various neurological diseases. Phase II activities will be focused on scaling up the manufacturing of neuronal EVs nanoprobes kit and developing high-throughput automatic sample processing system integrated with neuron- NPs kit.