Department of Neuroscience, Mayo Clinic, Jacksonville, Florida, United States of America.
Neurobiology of Disease Graduate Program, Mayo Graduate School, Mayo Clinic College of Medicine, Rochester, Minnesota, United States of America.
PLoS One. 2024 Apr 18;19(4):e0298080. doi: 10.1371/journal.pone.0298080. eCollection 2024.
Inclusions containing TAR DNA binding protein 43 (TDP-43) are a pathological hallmark of frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS). One of the disease-specific features of TDP-43 inclusions is the aberrant phosphorylation of TDP-43 at serines 409/410 (pS409/410). Here, we developed rabbit monoclonal antibodies (mAbs) that specifically detect pS409/410-TDP-43 in multiple model systems and FTD/ALS patient samples. Specifically, we identified three mAbs (26H10, 2E9 and 23A1) from spleen B cell clones that exhibit high specificity and sensitivity to pS409/410-TDP-43 peptides in an ELISA assay. Biochemical analyses revealed that pS409/410 of recombinant TDP-43 and of exogenous 25 kDa TDP-43 C-terminal fragments in cultured HEK293T cells are detected by all three mAbs. Moreover, the mAbs detect pS409/410-positive TDP-43 inclusions in the brains of FTD/ALS patients and mouse models of TDP-43 proteinopathy by immunohistochemistry. Our findings indicate that these mAbs are a valuable resource for investigating TDP-43 pathology both in vitro and in vivo.
包含 TAR DNA 结合蛋白 43（TDP-43）的包含物是额颞叶痴呆（FTD）和肌萎缩性侧索硬化症（ALS）的病理学标志之一。TDP-43 包含物中 TDP-43 的异常磷酸化是其疾病特异性特征之一，即在丝氨酸 409/410（pS409/410）处磷酸化。在这里，我们开发了兔单克隆抗体（mAbs），可特异性检测多种模型系统和 FTD/ALS 患者样本中的 pS409/410-TDP-43。具体来说，我们从脾 B 细胞克隆中鉴定出三个 mAbs（26H10、2E9 和 23A1），它们在 ELISA 测定中对 pS409/410-TDP-43 肽具有高特异性和灵敏度。生化分析表明，所有三种 mAbs 均可检测重组 TDP-43 的 pS409/410 和培养的 HEK293T 细胞中外源 25 kDa TDP-43 C 端片段中的 pS409/410。此外，这些 mAbs 通过免疫组织化学检测 FTD/ALS 患者和 TDP-43 蛋白病小鼠模型大脑中的 pS409/410 阳性 TDP-43 包含物。我们的发现表明，这些 mAbs 是研究 TDP-43 病理学的有价值的资源，无论是在体外还是体内。
