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

• Published in: FEBS letters 2024-04, Vol.598 (8), p.902-914
• Main Authors: Ohgita, Takashi, Sakai, Koto, Fukui, Nodoka, Namba, Norihiro, Nakano, Miyu, Kiguchi, Yuki, Morita, Izumi, Oyama, Hiroyuki, Yamaki, Kouya, Nagao, Kohjiro, Kobayashi, Norihiro, Saito, Hiroyuki
• Format: Article
• Language: English
• Subjects: amyloid; apoE isoform; apolipoprotein E; brain; cholesterol; dissociation; enzyme-linked immunosorbent assay; epitopes; humans; interferometry; lipid metabolism; monoclonal antibody; sandwich ELISA
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1002/1873-3468.14858

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. Apolipoprotein E (apoE) is a regulator of lipid metabolism, cholesterol transport, and the clearance and aggregation of amyloid β in the brain. Human apoE4 isoform is a risk factor for apoE‐related diseases, although only one or two residues are different in other isoforms. Here, we generated novel anti‐apoE monoclonal antibodies and constructed a sandwich ELISA system to selectively detect the apoE4 isoform.