Department of Neuroscience, College of Medicine, University of Florida, Gainesville, FL 32610, USA; Center for Translational Research in Neurodegenerative Disease, College of Medicine, University of Florida, Gainesville, FL 32610, USA.
Department of Neuroscience, College of Medicine, University of Florida, Gainesville, FL 32610, USA; Center for Translational Research in Neurodegenerative Disease, College of Medicine, University of Florida, Gainesville, FL 32610, USA; McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, FL 32610, USA.
Neurosci Lett. 2021 Jan 18;742:135553. doi: 10.1016/j.neulet.2020.135553. Epub 2020 Dec 17.
Amyotrophic lateral sclerosis (ALS) is a progressive neurological disease that leads to motor neuron degeneration and paralysis. Superoxide dismutase (SOD1) mutations are the second most common cause of familial ALS and are responsible for up to 20 % of familial ALS cases. In ALS patients, SOD1 can form toxic misfolded aggregates that deposit in the brain and spinal cord. To better detect SOD1 aggregates and expand the repertoire of conformational SOD1 antibodies, SOD1 monoclonal antibodies were generated by immunizing SOD1 knockout mice with an SOD1 fragment consisting of amino acids 129-146, which make up part of the electrostatic loop. A series of hybridomas secreting antibodies were screened and five different SOD1 monoclonal antibodies (2C10, 2F8, 4B11, 5H5, and 5A10) were found to preferentially detect denatured or aggregated SOD1 by enzyme-linked immunosorbent assay (ELISA), filter trap assay, and immunohistochemical analysis in SOD1 mouse models. The staining with these antibodies was compared to Campbell-Switzer argyrophilic reactivity of pathological inclusions. These new conformational selective SOD1 antibodies will be useful for clinical diagnosis of SOD1 ALS and potentially for passive immunotherapy.
肌萎缩侧索硬化症（ALS）是一种进行性神经疾病，导致运动神经元变性和瘫痪。超氧化物歧化酶 1（SOD1）突变是家族性 ALS 的第二大常见原因，占家族性 ALS 病例的 20%左右。在 ALS 患者中，SOD1 可以形成有毒的错误折叠聚集体，在大脑和脊髓中沉积。为了更好地检测 SOD1 聚集体并扩大构象 SOD1 抗体的库，通过用包含氨基酸 129-146 的 SOD1 片段免疫 SOD1 敲除小鼠，生成了 SOD1 单克隆抗体，该片段构成了部分静电环。筛选了一系列分泌抗体的杂交瘤，并发现了 5 种不同的 SOD1 单克隆抗体（2C10、2F8、4B11、5H5 和 5A10），通过酶联免疫吸附测定（ELISA）、滤膜陷阱测定和 SOD1 小鼠模型的免疫组织化学分析，这些抗体优先检测变性或聚集的 SOD1。这些新的构象选择性 SOD1 抗体将有助于 SOD1 ALS 的临床诊断，并可能有助于被动免疫治疗。
