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Immunological discrimination between the human apolipoprotein E2(Arg158----Cys) and E3 isoforms.
Authors:K Gerritse  P de Knijff  G van Ierssel  L M Havekes  R R Frants  M M Schellekens  N D Zegers  E Claassen  W J Boersma
Institution:Medical Biological Laboratory TNO, Rijswijk, The Netherlands.
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.
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