Open AccessConformation‐specific antibodies against multiple amyloid protofibril species from a single amyloid immunogen
Author(s) -
BonitoOliva Alessandra,
SchedinWeiss Sophia,
Younesi Shahab S.,
Tiiman Ann,
Adura Carolina,
Paknejad Navid,
Brendel Matt,
Romin Yevgeniy,
Parchem Ronald J.,
Graff Caroline,
Vukojević Vladana,
Tjernberg Lars O.,
Terenius Lars,
Winblad Bengt,
Sakmar Thomas P.,
Graham W Vallen
Publication year - 2019
Publication title -
journal of cellular and molecular medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.44
H-Index - 130
eISSN - 1582-4934
pISSN - 1582-1838
DOI - 10.1111/jcmm.14119
Subject(s) - immunogen , epitope , monoclonal antibody , amyloid (mycology) , chemistry , peptide , antibody , in vitro , microbiology and biotechnology , antigen , biochemistry , biology , immunology , inorganic chemistry
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 (Aβ42) protofibrils, but not with Aβ40 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 Aβ42, 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.