Parkinson’s disease (PD) is a progressive neurodegenerative disorder characterized by motor symptoms such as tremors, rigidity, and bradykinesia, as well as non-motor symptoms including cognitive impairment and mood ...Parkinson’s disease (PD) is a progressive neurodegenerative disorder characterized by motor symptoms such as tremors, rigidity, and bradykinesia, as well as non-motor symptoms including cognitive impairment and mood disorders. A hallmark of PD is the accumulation of alpha-synuclein, a presynaptic neuronal protein that aggregates to form Lewy bodies, leading to neuronal dysfunction and cell death. The study of alpha-synuclein and its pathological forms is crucial for understanding the etiology of PD and developing effective diagnostic and therapeutic strategies. Analytical techniques play a pivotal role in elucidating the structure, function, and aggregation mechanisms of alpha-synuclein. Biochemical methods such as Western blotting and enzyme-linked immunosorbent assay (ELISA) are employed to detect and quantify alpha-synuclein in biological samples, offering insights into its expression levels and post-translational modifications. Imaging techniques like immunohistochemistry and positron emission tomography (PET) allow for the visualization of alpha-synuclein aggregates in tissue samples and in vivo, respectively, facilitating the study of its spatial distribution and progression in PD Spectroscopic methods, including nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry, provide detailed structural information on alpha-synuclein and its isoforms, aiding in the identification of conformational changes associated with aggregation. Emerging techniques such as cryo-electron microscopy (Cryo-EM) and single-molecule fluorescence enable high-resolution structural analysis and real-time monitoring of alpha-synuclein aggregation dynamics, respectively. The application of these analytical techniques has significantly advanced our understanding of the pathophysiological role of alpha-synuclein in PD. They have contributed to the identification of potential biomarkers for early diagnosis and the evaluation of therapeutic interventions targeting alpha-synuclein aggregation. Despite technical limitations and challenges in clinical translation, ongoing advancements in analytical methodologies hold promise for improving the diagnosis, monitoring, and treatment of Parkinson’s disease through a deeper understanding of alpha-synuclein pathology.展开更多
目的分析5种识别α-突触核蛋白(α-synuclein,α-syn)N端结构域的单克隆抗体1C16、2B8、2P21、3O18和1J6的特异性,为帕金森病(Parkinson s disease,PD)的早期诊断和免疫治疗提供抗体支持。方法体外蛋白重组技术获得人源α-syn蛋白(human...目的分析5种识别α-突触核蛋白(α-synuclein,α-syn)N端结构域的单克隆抗体1C16、2B8、2P21、3O18和1J6的特异性,为帕金森病(Parkinson s disease,PD)的早期诊断和免疫治疗提供抗体支持。方法体外蛋白重组技术获得人源α-syn蛋白(human-α-syn,h-α-syn)、鼠源α-syn蛋白(mouse-α-syn,m-α-syn)、β-syn蛋白、N端人源α-syn蛋白(α-syn/N)和去N端人源α-syn蛋白(α-syn/ΔN)。斑点印迹法鉴定5种单克隆抗体的特异性识别结构域,使用蛋白质印迹技术检测其对变性后的纯蛋白和鼠脑组织的识别情况。结果抗体1C16可以识别非变性的h-α-syn纯蛋白和小鼠脑组织中的变性的h-α-syn蛋白,不识别m-α-syn和β-syn。抗体1J6仅识别非变性的h-α-syn及α-syn/N纯蛋白,不识别变性后的全长α-syn纯蛋白和小鼠脑组织中的变性h-α-syn蛋白。结论筛选出的2种单克隆抗体具有特异性,可为本实验下一步生物标志物的酶联免疫吸附法测定检测和针对α-syn的免疫治疗提供基础。展开更多
文摘Parkinson’s disease (PD) is a progressive neurodegenerative disorder characterized by motor symptoms such as tremors, rigidity, and bradykinesia, as well as non-motor symptoms including cognitive impairment and mood disorders. A hallmark of PD is the accumulation of alpha-synuclein, a presynaptic neuronal protein that aggregates to form Lewy bodies, leading to neuronal dysfunction and cell death. The study of alpha-synuclein and its pathological forms is crucial for understanding the etiology of PD and developing effective diagnostic and therapeutic strategies. Analytical techniques play a pivotal role in elucidating the structure, function, and aggregation mechanisms of alpha-synuclein. Biochemical methods such as Western blotting and enzyme-linked immunosorbent assay (ELISA) are employed to detect and quantify alpha-synuclein in biological samples, offering insights into its expression levels and post-translational modifications. Imaging techniques like immunohistochemistry and positron emission tomography (PET) allow for the visualization of alpha-synuclein aggregates in tissue samples and in vivo, respectively, facilitating the study of its spatial distribution and progression in PD Spectroscopic methods, including nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry, provide detailed structural information on alpha-synuclein and its isoforms, aiding in the identification of conformational changes associated with aggregation. Emerging techniques such as cryo-electron microscopy (Cryo-EM) and single-molecule fluorescence enable high-resolution structural analysis and real-time monitoring of alpha-synuclein aggregation dynamics, respectively. The application of these analytical techniques has significantly advanced our understanding of the pathophysiological role of alpha-synuclein in PD. They have contributed to the identification of potential biomarkers for early diagnosis and the evaluation of therapeutic interventions targeting alpha-synuclein aggregation. Despite technical limitations and challenges in clinical translation, ongoing advancements in analytical methodologies hold promise for improving the diagnosis, monitoring, and treatment of Parkinson’s disease through a deeper understanding of alpha-synuclein pathology.
文摘目的分析5种识别α-突触核蛋白(α-synuclein,α-syn)N端结构域的单克隆抗体1C16、2B8、2P21、3O18和1J6的特异性,为帕金森病(Parkinson s disease,PD)的早期诊断和免疫治疗提供抗体支持。方法体外蛋白重组技术获得人源α-syn蛋白(human-α-syn,h-α-syn)、鼠源α-syn蛋白(mouse-α-syn,m-α-syn)、β-syn蛋白、N端人源α-syn蛋白(α-syn/N)和去N端人源α-syn蛋白(α-syn/ΔN)。斑点印迹法鉴定5种单克隆抗体的特异性识别结构域,使用蛋白质印迹技术检测其对变性后的纯蛋白和鼠脑组织的识别情况。结果抗体1C16可以识别非变性的h-α-syn纯蛋白和小鼠脑组织中的变性的h-α-syn蛋白,不识别m-α-syn和β-syn。抗体1J6仅识别非变性的h-α-syn及α-syn/N纯蛋白,不识别变性后的全长α-syn纯蛋白和小鼠脑组织中的变性h-α-syn蛋白。结论筛选出的2种单克隆抗体具有特异性,可为本实验下一步生物标志物的酶联免疫吸附法测定检测和针对α-syn的免疫治疗提供基础。
