Reactivity with erythroid and non-erythroid tissues of a murine monoclonal antibody to a synthetic peptide having amino acid sequence common to cytoplasmic domain of human glycophorins C and D.
Three synthetic peptides encompassing the entire cytoplasmic polypeptide sequence (amino acid residues 82-128) of glycophorin C (GPC) and glycophorin D (GPD) were used to immunize mice for the production of monoclonal antibodies (MoAbs). Only the synthetic peptide (GPC-peptide-1) corresponding to C-terminal residues 112-128 elicited a MoAb (named BGRL-100) which could react with native and denatured GPC and GPD. We characterized BGRL-100 by inhibition using GPC-peptide 1 and red cell sialoglycoproteins. The ability of BGRL-100 to interact with native GPC and GPD was assessed by immunoprecipitation with normal red cells (RBCs), and with denatured GPC and GPD by Western blotting of both normal RBCs and RBCs carrying GPC variants. Immunohistochemical staining of human tissue sections was performed using both BGRL-100 and a rat MoAb (named BRAC-1), which is specific for an extracellular domain of GPC and GPD. Both antibodies showed strong staining of erythroid lineage haemopoietic cells in fetal liver, sinusoids of adult liver and RBCs in the blood vessels of all tissues tested. Neither antibody reacted with epithelia from a range of human tissues. However, both MoAbs stained neural tissue in a distinctive fibrillar pattern. This suggests the presence of an analogue of erythroid GPC in neural tissues.
King
M J
MJ
International Blood Group Reference Laboratory, Bristol, U.K.
Holmes
C H
CH
Mushens
R E
RE
Mawby
W
W
Reid
M E
ME
Scott
M L
ML
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D001667:Binding, Competitive; D015153:Blotting, Western; D004912:Erythrocytes / Q000276:immunology*; D006021:Glycophorins / Q000032:analysis / Q000276:immunology*; D006412:Hematopoietic Stem Cells / Q000737:chemistry; D006801:Humans; D008099:Liver / Q000737:chemistry / Q000196:embryology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000276:immunology*; D011233:Precipitin Tests; D051381:Rats
D000911:Antibodies, Monoclonal; D006021:Glycophorins; D010446:Peptide Fragments