Immunological discrimination between the human apolipoprotein E2(Arg158----Cys) and E3 isoforms.
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.
Gerritse
K
K
Medical Biological Laboratory TNO, Rijswijk, The Netherlands.
de Knijff
P
P
van Ierssel
G
G
Havekes
L M
LM
Frants
R R
RR
Schellekens
M M
MM
Zegers
N D
ND
Claassen
E
E
Boersma
W J
WJ
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
D000911:Antibodies, Monoclonal; D053329:Apolipoprotein E2; D053318:Apolipoprotein E3; D001057:Apolipoproteins E; D000939:Epitopes; D001120:Arginine; D003545:Cysteine