Identification of an epitope recognized by the monoclonal antibody PEP80 in the C-terminal cytoplasmic fragment of glycophorin A.
The monoclonal antibody PEP80 (IgG1) was raised by immunization of BALB/c mice with asialo-agalacto-glycophorin from human erythrocytes. The antibody is specific for glycophorin A (GPA) and reacts strongly with the GPA-derived tryptic peptide which is the C-terminal cytoplasmic portion of GPA, containing amino acid residues 102-131. Using the smaller chymotryptic fragments of this peptide and a set of solid phase-synthesized peptides allowed to establish that the MAb PEP80 is directed against an epitope comprising amino acid residues 112-121 of GPA. The peptides terminated with 120th or 119th amino acid residue were slightly less active, and the minimal structure which still gave a weak reaction with the antibody was the sequence of amino acid residues 112-118. The MAb PEP80 did not bind to live human erythroleukemic K562 cells, but showed a strong binding to the cells permeabilized with methanol.
Duk
M
M
Department of Immunochemistry, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wroclaw.
Czerwiński
M
M
Lisowska
E
E
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D003593:Cytoplasm / Q000276:immunology; D000939:Epitopes / Q000737:chemistry; D006021:Glycophorins / Q000737:chemistry / Q000276:immunology*; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000939:Epitopes; D006021:Glycophorins; D010446:Peptide Fragments