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Novel conformation‐selective monoclonal antibodies against apoA‐I amyloid fibrils.

Authors :
Ohgita, Takashi
Furutani, Yuki
Nakano, Miyu
Hattori, Megumi
Suzuki, Ayane
Nakagawa, Miho
Naniwa, Sera
Morita, Izumi
Oyama, Hiroyuki
Nishitsuji, Kazuchika
Kobayashi, Norihiro
Saito, Hiroyuki
Source :
FEBS Journal; Mar2021, Vol. 288 Issue 5, p1496-1513, 18p
Publication Year :
2021

Abstract

The Iowa (G26R) mutation in human apolipoprotein A‐I (apoA‐I), the major protein of plasma high‐density lipoprotein, is associated with systemic amyloidosis, and the N‐terminal 1–83 fragment of apoA‐I carrying this mutation has a strong propensity to form amyloid fibrils. Here, we generated and characterized novel monoclonal antibodies (mAbs) that display selective reactivity to apoA‐I amyloid fibrils. By immunizing BALB/c and A/J mice with apoA‐I 1‒83/G26R fibrils conjugated with hemocyanin and the hybridoma production, four IgM class mAbs were obtained. The generated mAbs exhibited strong reactivity to amyloid fibrils formed by the 1–83 fragment of apoA‐I, but not to the monomeric 1–83 fragment or full‐length apoA‐I. The apparent dissociation constant of the mAbs to apoA‐I fibrils was determined to be within the nM range. A time‐dependent aggregation assay demonstrated that the mAbs preferentially react with mature fibrils over non‐fibrillar aggregates formed by apoA‐I 1–83/G26R. In addition, dot blotting and ELISA using deletion or proline substituted variants of apoA‐I 1‒83/G26R suggest that the generated mAbs react to common structural features in apoA‐I amyloid fibrils. Indeed, the mAbs also recognized amyloid fibrils formed by α‐synuclein that has no sequence identity to apoA‐I. Thus, our newly generated anti‐apoA‐I fibril mAbs may be utilized for not only diagnosis of apoA‐I‐related amyloidosis but also structural analysis of amyloid fibrils as novel conformation‐selective antibodies. [ABSTRACT FROM AUTHOR]

Details

Language :
English
ISSN :
1742464X
Volume :
288
Issue :
5
Database :
Complementary Index
Journal :
FEBS Journal
Publication Type :
Academic Journal
Accession number :
148995942
Full Text :
https://doi.org/10.1111/febs.15487