Generation and characterization of novel conformation-specific monoclonal antibodies for α-synuclein pathology.
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.
Department of Biochemistry, College of Medicine and Health Science, United Arab Emirates University, Al Ain, United Arab Emirates; Neural Plasticity and Repair Unit, Department of Experimental Medical Sciences, Wallenberg Neuroscience Center, BMC A10, Lund University, Lund, Sweden.
Nishant N
NN
Vaikath
Department of Biochemistry, College of Medicine and Health Science, United Arab Emirates University, Al Ain, United Arab Emirates; Department of Anatomy and Neurosciences, Neuroscience Campus Amsterdam, VU University Medical Center, Amsterdam, The Netherlands.
Nour K
NK
Majbour
Department of Molecular Biology and Genetics, Democritus University of Thrace, Alexandroupolis, Greece.
Katerina E
KE
Paleologou
Department of Biochemistry, College of Medicine and Health Science, United Arab Emirates University, Al Ain, United Arab Emirates.
Mustafa T
MT
Ardah
Department of Anatomy and Neurosciences, Neuroscience Campus Amsterdam, VU University Medical Center, Amsterdam, The Netherlands.
Esther
E
van Dam
Department of Anatomy and Neurosciences, Neuroscience Campus Amsterdam, VU University Medical Center, Amsterdam, The Netherlands.
Wilma D J
WD
van de Berg
Discipline of Pathology, Charles Perkin Centre, University of Sydney, Sydney, Australia.
Shelley L
SL
Forrest
Department of Clinical Neurology, University of Oxford, Oxford, UK.
Laura
L
Parkkinen
Department of Human Physiology, School of Medicine, Flinders University, Australia.
Wei-Ping
WP
Gai
Department of Research for Parkinson's Disease, Juntendo University Graduate School of Medicine, Japan; Department of Neurology, Juntendo University Graduate School of Medicine, Japan.
Nobutaka
N
Hattori
Department of Neurology, Juntendo University Graduate School of Medicine, Japan.
Masashi
M
Takanashi
Neuroscience Research Institute, Department of Medicine, Seoul National University College of Medicine, Seoul 110-799, Korea.
Seung-Jae
SJ
Lee
Clinical and Cognitive Neuroscience Research Group, University of Manchester, Salford Royal Foundation NHS Trust, Salford M6 8HD, UK.
David M A
DM
Mann
Department of Research for Parkinson's Disease, Juntendo University Graduate School of Medicine, Japan.
Yuzuru
Y
Imai
Faculty of Medicine, University of New South Wales and Neuroscience Research Australia, Sydney, Australia.
Glenda M
GM
Halliday
Neural Plasticity and Repair Unit, Department of Experimental Medical Sciences, Wallenberg Neuroscience Center, BMC A10, Lund University, Lund, Sweden.
Jia-Yi
JY
Li
Department of Biochemistry, College of Medicine and Health Science, United Arab Emirates University, Al Ain, United Arab Emirates; College of Science, Engineering and Technology, HBKU, Education City, Qatar Foundation, Doha, Qatar. Electronic address: oelagnaf@qf.org.qa.
Omar M A
OM
El-Agnaf
D048868:Adaptor Proteins, Signal Transducing; D000544:Alzheimer Disease / Q000378:metabolism; D016229:Amyloid beta-Peptides / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D001921:Brain / Q000378:metabolism* / Q000473:pathology; D004926:Escherichia coli; D058228:Islet Amyloid Polypeptide / Q000378:metabolism; D020961:Lewy Body Disease / Q000378:metabolism / Q000473:pathology; D008562:Membrane Glycoproteins / Q000378:metabolism; D051379:Mice; D009363:Neoplasm Proteins / Q000276:immunology / Q000378:metabolism; D010300:Parkinson Disease / Q000378:metabolism / Q000473:pathology; D010446:Peptide Fragments / Q000378:metabolism; D011487:Protein Conformation; D055503:Protein Multimerization; D011994:Recombinant Proteins / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D051844:alpha-Synuclein / Q000737:chemistry* / Q000276:immunology* / Q000378:metabolism; D051846:beta-Synuclein / Q000276:immunology / Q000378:metabolism; D051845:gamma-Synuclein / Q000276:immunology / Q000378:metabolism; D016875:tau Proteins / Q000378:metabolism
D048868:Adaptor Proteins, Signal Transducing; D016229:Amyloid beta-Peptides; D000911:Antibodies, Monoclonal; C120083:ITM2B protein, human; D058228:Islet Amyloid Polypeptide; C054369:MAPT protein, human; D008562:Membrane Glycoproteins; D009363:Neoplasm Proteins; D010446:Peptide Fragments; D011994:Recombinant Proteins; C497604:SNCA protein, human; C497607:SNCB protein, human; C104249:SNCG protein, human; D051844:alpha-Synuclein; C075222:amyloid beta-protein (1-42); D051846:beta-Synuclein; D051845:gamma-Synuclein; D016875:tau Proteins
Conformation-specific monoclonal antibodies; Dementia with Lewy bodies; Parkinson’s disease; α-Synuclein