Generation and characterization of novel conformation-specific monoclonal antibodies for alpha-synuclein pathology

Nishant Vaikath, Nour Majbour, K Paleologou, Mustafa Ardah, Esther van Dam, Wilma van de Berg, Shelley Forrest, Laura Parkkinen, Wei-Ping Gai, N Hattori, Masashi Takanashi, Seung-Jae Lee, David Mann, Yuzuru Imai, Glenda Halliday, Jia-Yi Li, Omar El-Agnaf

    Research output: Contribution to journalArticle

    55 Citations (Scopus)

    Abstract

    α-Synuclein (α-syn), a small protein that has the intrinsic propensity to aggregate, is implicated in several neurodegenerative diseases including Parkinson's disease (PD), dementia with Lewy bodies (DLB), and multiple system atrophy (MSA), which are collectively known as synucleinopathies. Genetic, pathological, biochemical, and animal modeling studies provided compelling evidence that α-syn aggregation plays a key role in the pathogenesis of PD and related synucleinopathies. It is therefore of utmost importance to develop reliable tools that can detect the aggregated forms of α-syn. We describe here the generation and characterization of six novel conformation-specific monoclonal antibodies that recognize specifically α-syn aggregates but not the soluble, monomeric form of the protein. The antibodies described herein did not recognize monomers or fibrils generated from other amyloidogenic proteins including β-syn, γ-syn, β-amyloid, tau protein, islet amyloid polypeptide and ABri. Interestingly, the antibodies did not react to overlapping linear peptides spanning the entire sequence of α-syn, confirming further that they only detect α-syn aggregates. In immunohistochemical studies, the new conformation-specific monoclonal antibodies showed underappreciated small micro-aggregates and very thin neurites in PD and DLB cases that were not observed with generic pan antibodies that recognize linear epitope. Furthermore, employing one of our conformation-specific antibodies in a sandwich based ELISA, we observed an increase in levels of α-syn oligomers in brain lysates from DLB compared to Alzheimer's disease and control samples. Therefore, the conformation-specific antibodies portrayed herein represent useful tools for research, biomarkers development, diagnosis and even immunotherapy for PD and related pathologies.

    Original languageEnglish
    Pages (from-to)81-99
    Number of pages19
    JournalNeurobiology of Disease
    Volume79
    DOIs
    Publication statusPublished - 2015

    Fingerprint Dive into the research topics of 'Generation and characterization of novel conformation-specific monoclonal antibodies for alpha-synuclein pathology'. Together they form a unique fingerprint.

  • Cite this

    Vaikath, N., Majbour, N., Paleologou, K., Ardah, M., van Dam, E., van de Berg, W., Forrest, S., Parkkinen, L., Gai, W-P., Hattori, N., Takanashi, M., Lee, S-J., Mann, D., Imai, Y., Halliday, G., Li, J-Y., & El-Agnaf, O. (2015). Generation and characterization of novel conformation-specific monoclonal antibodies for alpha-synuclein pathology. Neurobiology of Disease, 79, 81-99. https://doi.org/10.1016/j.nbd.2015.04.009