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

FEBS Lett. 2024 Apr;598(8):902-914. doi: 10.1002/1873-3468.14858. Epub 2024 Mar 26.

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.

Keywords: apoE isoform; monoclonal antibody; sandwich ELISA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Monoclonal* / chemistry
  • Antibodies, Monoclonal* / immunology
  • Apolipoprotein E3 / chemistry
  • Apolipoprotein E3 / genetics
  • Apolipoprotein E3 / immunology
  • Apolipoprotein E3 / metabolism
  • Apolipoprotein E4 / genetics
  • Apolipoprotein E4 / immunology
  • Apolipoprotein E4 / metabolism
  • Apolipoproteins E* / chemistry
  • Apolipoproteins E* / genetics
  • Apolipoproteins E* / immunology
  • Apolipoproteins E* / metabolism
  • Enzyme-Linked Immunosorbent Assay / methods
  • Epitopes / chemistry
  • Epitopes / immunology
  • Humans
  • Mice
  • Mice, Inbred BALB C
  • Protein Isoforms* / immunology

Substances

  • Antibodies, Monoclonal
  • Protein Isoforms
  • Apolipoproteins E
  • Epitopes
  • Apolipoprotein E4
  • Apolipoprotein E3