title: Immunological discrimination between the human apolipoprotein E2(Arg158----Cys) and E3 isoforms.
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.
authors.affiliation: Medical Biological Laboratory TNO, Rijswijk, The Netherlands.
authors.forename: K
authors.initials: K
authors.lastname: Gerritse
authors.forename: P
authors.initials: P
authors.lastname: de Knijff
authors.forename: G
authors.initials: G
authors.lastname: van Ierssel
authors.forename: L M
authors.initials: LM
authors.lastname: Havekes
authors.forename: R R
authors.initials: RR
authors.lastname: Frants
authors.forename: M M
authors.initials: MM
authors.lastname: Schellekens
authors.forename: N D
authors.initials: ND
authors.lastname: Zegers
authors.forename: E
authors.initials: E
authors.lastname: Claassen
authors.forename: W J
authors.initials: WJ
authors.lastname: Boersma
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D053329:Apolipoprotein E2; D053318:Apolipoprotein E3; D001057:Apolipoproteins E / Q000737:chemistry / Q000276:immunology*; D001120:Arginine; D003545:Cysteine; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D005260:Female; D006801:Humans; D015151:Immunoblotting / Q000379:methods; D007525:Isoelectric Focusing; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010641:Phenotype; D016415:Sequence Alignment
chemical_list: D000911:Antibodies, Monoclonal; D053329:Apolipoprotein E2; D053318:Apolipoprotein E3; D001057:Apolipoproteins E; D000939:Epitopes; D001120:Arginine; D003545:Cysteine