Conformation‐specific antibodies against multiple amyloid protofibril species from a single amyloid immunogen

• Published in: Journal of cellular and molecular medicine Vol. 23; no. 3; pp. 2103 - 2114
• Main Authors: Bonito‐Oliva, Alessandra, Schedin‐Weiss, 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
• Format: Journal Article
• Language: English
• Published: England John Wiley & Sons, Inc 01.03.2019; John Wiley and Sons Inc
• Subjects: Alzheimer Disease - immunology; Alzheimer Disease - metabolism; Alzheimer's disease; Amylin; amyloid; Amyloid - immunology; Amyloid - metabolism; amyloid aggregation; Amyloid beta-Peptides - immunology; Amyloid beta-Peptides - metabolism; Animals; Antibodies, Monoclonal - chemistry; Antibodies, Monoclonal - immunology; Antibodies, Monoclonal - metabolism; Antibody Specificity - immunology; Brain - immunology; Brain - metabolism; Brain - pathology; chaperone‐like amyloid‐binding protein; conformation‐sensitive; Epitopes; Epitopes - chemistry; Epitopes - immunology; Epitopes - metabolism; Humans; Islet Amyloid Polypeptide - immunology; Islet Amyloid Polypeptide - metabolism; Medicin och hälsovetenskap; Mice; Monoclonal antibodies; monoclonal antibody; Monomers; Neurodegenerative diseases; Nucleobindins - immunology; Nucleobindins - metabolism; Original; Peptide Fragments - immunology; Peptide Fragments - metabolism; Protein Binding; Protein Conformation; protofibril; Pyramidal Cells - immunology; Pyramidal Cells - metabolism; Senile plaques; Tissues; β-Amyloid; protofibril; conformation-sensitive; amyloid; Alzheimer’s disease; amyloid aggregation; chaperone-like amyloid-binding protein; monoclonal antibody
• ISSN: 1582-1838; 1582-4934; 1582-4934
• DOI: 10.1111/jcmm.14119

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.