Generation of novel anti‐apoE monoclonal antibodies that selectively recognize apoE isoforms

Author:

Ohgita Takashi12ORCID,Sakai Koto1,Fukui Nodoka1,Namba Norihiro1,Nakano Miyu1,Kiguchi Yuki3,Morita Izumi3,Oyama Hiroyuki3,Yamaki Kouya4,Nagao Kohjiro1,Kobayashi Norihiro3,Saito Hiroyuki1ORCID

Affiliation:

1. Laboratory of Biophysical Chemistry Kyoto Pharmaceutical University Japan

2. Center for Instrumental Analysis Kyoto Pharmaceutical University Japan

3. Laboratory of Bioanalytical Chemistry Kobe Pharmaceutical University Japan

4. Laboratory of Pharmacology Kobe Pharmaceutical University Japan

Abstract

Apolipoprotein E (apoE) is a regulator of lipid metabolism, cholesterol transport, and the clearance and aggregation of amyloid β in the brain. The three human apoE isoforms apoE2, apoE3, and apoE4 only differ in one or two residues. Nevertheless, the functions highly depend on the isoform types and lipidated states. Here, we generated novel anti‐apoE monoclonal antibodies (mAbs) and obtained an apoE4‐selective mAb whose epitope is within residues 110–117. ELISA and bio‐layer interferometry measurements demonstrated that the dissociation constants of mAbs are within the nanomolar range. Using the generated antibodies, we successfully constructed sandwich ELISA systems, which can detect all apoE isoforms or selectively detect apoE4. These results suggest the usability of the generated anti‐apoE mAbs for selective detection of apoE isoforms.

Funder

Kyoto Pharmaceutical University

Japan Society for the Promotion of Science

Japan Private School Promotion Foundation

Publisher

Wiley

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