Repository hosted by TU Delft Library

Home · Contact · About · Disclaimer ·
 

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

Author: Gerritse, K. · Knijff, P. de · Ierssel, G.V. · Havekes, L.M. · Frants, R.R. · Schellekens, M.M. · Zegers, N.D. · Claassen, E. · Boersma, W.J.A.
Type:article
Date:1992
Institution: Med. Biological Laboratory-TNO, Immunol./Med. Microbiol. Dept. Instituut voor verouderings- en vaatziekten onderzoek TNO
Source:Journal of Lipid Research, 2, 33, 273-280
Identifier: 231803
Keywords: Synthetic peptides · Type III hyperlipoproteinemia · Apolipoprotein e2 · Apolipoprotein e3 · Lipoprotein receptor · Monoclonal antibody · Amino acid sequence · Antigenicity · Binding affinity · Immunological tolerance · Lipid metabolism · Lipoprotein blood level · Nonhuman · Sequence homology · Amino Acid Sequence · Animal · Antibodies, Monoclonal · Antibody Specificity · Apolipoproteins E · Arginine · Cysteine · Enzyme-Linked Immunosorbent Assay · Epitopes · Female · Human · Immunoblotting · Isoelectric Focusing · Mice · Mice, Inbred BALB C · Molecular Sequence Data · Phenotype · Sequence Alignment

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. Chemicals/CAS: arginine, 1119-34-2, 15595-35-4, 7004-12-8, 74-79-3; cysteine, 4371-52-2, 52-89-1, 52-90-4; Antibodies, Monoclonal; apolipoprotein E-2; apolipoprotein E-3; Apolipoproteins E; Arginine, 74-79-3; Cysteine, 52-90-4; Epitopes