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Rational design of a conformation-specific antibody for the quantification of Aβ oligomers.

Francesco Antonio AprilePietro SormanniMarina PodpolnyShianne ChhangurLisa-Maria NeedhamFrancesco Simone RuggeriMichele PerniRyan LimbockerGabriella T HellerTomas SneiderisTom ScheidtBenedetta ManniniJohnny HabchiSteven F LeePatricia C SalinasTuomas P J KnowlesChristopher M DobsonChristopher M Dobson
Published in: Proceedings of the National Academy of Sciences of the United States of America (2020)
Protein misfolding and aggregation is the hallmark of numerous human disorders, including Alzheimer's disease. This process involves the formation of transient and heterogeneous soluble oligomers, some of which are highly cytotoxic. A major challenge for the development of effective diagnostic and therapeutic tools is thus the detection and quantification of these elusive oligomers. Here, to address this problem, we develop a two-step rational design method for the discovery of oligomer-specific antibodies. The first step consists of an "antigen scanning" phase in which an initial panel of antibodies is designed to bind different epitopes covering the entire sequence of a target protein. This procedure enables the determination through in vitro assays of the regions exposed in the oligomers but not in the fibrillar deposits. The second step involves an "epitope mining" phase, in which a second panel of antibodies is designed to specifically target the regions identified during the scanning step. We illustrate this method in the case of the amyloid β (Aβ) peptide, whose oligomers are associated with Alzheimer's disease. Our results show that this approach enables the accurate detection and quantification of Aβ oligomers in vitro, and in Caenorhabditis elegans and mouse hippocampal tissues.
Keyphrases
  • cognitive decline
  • endothelial cells
  • high throughput
  • amino acid
  • gene expression
  • loop mediated isothermal amplification
  • cerebral ischemia
  • real time pcr
  • binding protein
  • brain injury
  • blood brain barrier