Conformation-specific antibodies against multiple amyloid protofibril species from a single amyloid immunogen.
J Cell Mol Med
; 23(3): 2103-2114, 2019 03.
Article
de En
| MEDLINE
| ID: mdl-30663210
ABSTRACT
We engineered and employed a chaperone-like amyloid-binding protein Nucleobindin 1 (NUCB1) to stabilize human islet amyloid polypeptide (hIAPP) protofibrils for use as immunogen in mice. We obtained multiple monoclonal antibody (mAb) clones that were reactive against hIAPP protofibrils. A secondary screen was carried out to identify clones that cross-reacted with amyloid beta-peptide (Aß42) protofibrils, but not with Aß40 monomers. These mAbs were further characterized in several in vitro assays, in immunohistological studies of a mouse model of Alzheimer's disease (AD) and in AD patient brain tissue. We show that mAbs obtained by immunizing mice with the NUCB1-hIAPP complex cross-react with Aß42, specifically targeting protofibrils and inhibiting their further aggregation. In line with conformation-specific binding, the mAbs appear to react with an intracellular antigen in diseased tissue, but not with amyloid plaques. We hypothesize that the mAbs we describe here recognize a secondary or quaternary structural epitope that is common to multiple amyloid protofibrils. In summary, we report a method to create mAbs that are conformation-sensitive and sequence-independent and can target more than one type of protofibril species.
Mots clés
Texte intégral:
1
Collection:
01-internacional
Base de données:
MEDLINE
Sujet principal:
Fragments peptidiques
/
Peptides bêta-amyloïdes
/
Amyloïde
/
Anticorps monoclonaux
Limites:
Animals
/
Humans
Langue:
En
Journal:
J Cell Mol Med
Sujet du journal:
BIOLOGIA MOLECULAR
Année:
2019
Type de document:
Article