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J Biol Chem. 2015 Feb 13;290(7):4059-74. doi: 10.1074/jbc.M114.627919. Epub 2014 Dec 24.

Tau monoclonal antibody generation based on humanized yeast models: impact on Tau oligomerization and diagnostics.

Author information

1
From Functional Biology, KU Leuven, Kasteelpark Arenberg 31 Box 2433, 3001 Heverlee, Belgium.
2
From Functional Biology, KU Leuven, Kasteelpark Arenberg 31 Box 2433, 3001 Heverlee, Belgium, ADx NeuroSciences NV, Technologiepark Zwijnaarde 4, 9052 Ghent, Belgium, Fujirebio Europe, Technologiepark Zwijnaarde 6, 9052 Ghent, Belgium.
3
INSERM, UMR1172, JPArc, Alzheimer & Tauopathies, Rue Polonovski, 59045 Lille, France, the Faculté de Médecine, Université de Lille, Place de Verdun, 59045 Lille, France, the Memory Clinic, Centre Hospitalier Régional Universitaire de Lille, 59037 Lille, France.
4
ADx NeuroSciences NV, Technologiepark Zwijnaarde 4, 9052 Ghent, Belgium.
5
Fujirebio Europe, Technologiepark Zwijnaarde 6, 9052 Ghent, Belgium.
6
Université Lille Nord de France, 59000 Lille, France, CNRS, UMR8576 Structural and Functional Glycobiology, 59650 Villeneuve d'Ascq, France.
7
the Reference Center for Biological Markers of Dementia (BIODEM), Institute Born-Bunge, University of Antwerp, 2610 Wilrijk, Belgium, and the Department of Neurology and Memory Clinic, Hospital Network Antwerp (ZNA) Middelheim and Hoge Beuken, 2660 Antwerp, Belgium.
8
ADx NeuroSciences NV, Technologiepark Zwijnaarde 4, 9052 Ghent, Belgium, Eugeen.vanmechelen@adxneurosciences.com.
9
From Functional Biology, KU Leuven, Kasteelpark Arenberg 31 Box 2433, 3001 Heverlee, Belgium, joris.winderickx@bio.kuleuven.be.

Abstract

A link between Tau phosphorylation and aggregation has been shown in different models for Alzheimer disease, including yeast. We used human Tau purified from yeast models to generate new monoclonal antibodies, of which three were further characterized. The first antibody, ADx201, binds the Tau proline-rich region independently of the phosphorylation status, whereas the second, ADx215, detects an epitope formed by the Tau N terminus when Tau is not phosphorylated at Tyr(18). For the third antibody, ADx210, the binding site could not be determined because its epitope is probably conformational. All three antibodies stained tangle-like structures in different brain sections of THY-Tau22 transgenic mice and Alzheimer patients, and ADx201 and ADx210 also detected neuritic plaques in the cortex of the patient brains. In hippocampal homogenates from THY-Tau22 mice and cortex homogenates obtained from Alzheimer patients, ADx215 consistently stained specific low order Tau oligomers in diseased brain, which in size correspond to Tau dimers. ADx201 and ADx210 additionally reacted to higher order Tau oligomers and presumed prefibrillar structures in the patient samples. Our data further suggest that formation of the low order Tau oligomers marks an early disease stage that is initiated by Tau phosphorylation at N-terminal sites. Formation of higher order oligomers appears to require additional phosphorylation in the C terminus of Tau. When used to assess Tau levels in human cerebrospinal fluid, the antibodies permitted us to discriminate patients with Alzheimer disease or other dementia like vascular dementia, indicative that these antibodies hold promising diagnostic potential.

KEYWORDS:

Alzheimer Disease; Antibody; Protein Folding; Protein Oligomer; Tau Protein (Tau); Yeast

PMID:
25540200
PMCID:
PMC4326816
DOI:
10.1074/jbc.M114.627919
[Indexed for MEDLINE]
Free PMC Article

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