Immunological discrimination between the human apolipoprotein E2(Arg158----Cys) and E3 isoforms

J Lipid Res. 1992 Feb;33(2):273-80.

Abstract

A specific anti-apoE2(Arg158----Cys) monoclonal antibody was raised by means of immunization of mice with a variant specific synthetic peptide. The peptide sequences used were homologous to apolipoprotein E of human and mouse. Consequently, the mouse immune system was tolerant to most of the selected sequences. Immunization with only one of selected peptides (amino acids 154-172) evoked an anti-peptide and anti-native protein response. Surprisingly, this peptide was predicted to have a low antigenicity index, in contrast to the other used peptides. The variant specific anti-peptide MAb that was generated with this sequence, recognizes apoE2(Arg158----Cys) and not apoE3. We here describe a sensitive, time saving, and simple immunoblot assay to detect apoE2(Arg158----Cys) in human sera without prior isoelectric focusing of serum proteins.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies, Monoclonal / immunology*
  • Antibody Specificity
  • Apolipoprotein E2
  • Apolipoprotein E3
  • Apolipoproteins E / chemistry
  • Apolipoproteins E / immunology*
  • Arginine
  • Cysteine
  • Enzyme-Linked Immunosorbent Assay
  • Epitopes
  • Female
  • Humans
  • Immunoblotting / methods
  • Isoelectric Focusing
  • Mice
  • Mice, Inbred BALB C
  • Molecular Sequence Data
  • Phenotype
  • Sequence Alignment

Substances

  • Antibodies, Monoclonal
  • Apolipoprotein E2
  • Apolipoprotein E3
  • Apolipoproteins E
  • Epitopes
  • Arginine
  • Cysteine