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

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

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alzheimer Disease / cerebrospinal fluid
  • Alzheimer Disease / diagnosis*
  • Alzheimer Disease / immunology
  • Animals
  • Antibodies, Monoclonal*
  • Biotinylation
  • Blotting, Western
  • Brain / immunology
  • Brain / metabolism
  • Brain / pathology*
  • Disease Progression
  • Enzyme-Linked Immunosorbent Assay
  • Epitope Mapping
  • Hippocampus / immunology
  • Hippocampus / metabolism
  • Hippocampus / pathology*
  • Humans
  • Immunization
  • Immunoenzyme Techniques
  • Immunoprecipitation
  • Magnetic Resonance Spectroscopy
  • Membrane Microdomains
  • Mice
  • Mice, Inbred BALB C
  • Mice, Transgenic
  • Neurofibrillary Tangles
  • Peptide Fragments / metabolism
  • Phosphorylation
  • Plaque, Amyloid
  • Saccharomyces cerevisiae
  • tau Proteins / cerebrospinal fluid
  • tau Proteins / chemistry*
  • tau Proteins / immunology*

Substances

  • Antibodies, Monoclonal
  • Peptide Fragments
  • tau Proteins