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The murine monoclonal antibody NaM26‐4C6 identifies a common structure on band 3 and glycophorin C
Author(s) -
M.J. Loirat,
Marcin Czerwiński,
Maria Duk,
Dominique Blanchard
Publication year - 1999
Publication title -
transfusion medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.471
H-Index - 59
eISSN - 1365-3148
pISSN - 0958-7578
DOI - 10.1046/j.1365-3148.1999.009001069.x
Subject(s) - glycophorin , monoclonal antibody , antibody , immunology , medicine , antigen
The murine monoclonal antibody NaM26‐4C6 (IgM class), obtained from the splenocytes of a BALB/c mouse immunized with human umbilical cord red blood cells, was characterized by agglutination test and immunoblotting analysis. The structure of the NaM26‐4C6 epitope was further elucidated by using a series of peptides synthesized on pins. The antibody agglutinated untreated and chymotrypsin‐treated but not trypsin‐ or neuraminidase‐treated human erythrocytes. Agglutination‐inhibition test demonstrated that the antibody recognizes an epitope located on the N‐terminal trypsin‐sensitive portion of glycophorin C. The antibody bound on immunoblots to glycophorin C, and also to the band 3 protein and its 69‐kDa N‐terminal fragment but did not bind to desialylated and de‐O‐glycosylated glycophorin C. Peptide mapping allowed localization of the binding site on the 23‐kDa N‐terminal intracellular peptide of band 3. The antibody binds to the amino‐acid sequences 22 EDPDIP 27 of band 3 protein and 15 SLEPDPGM 22 of glycophorin C, and residues D and P were found to be essential. The new epitope identified by NaM26‐4C6 corresponds to a linear amino acid sequence located on the N‐terminal intracellular portion of band 3 and to a more complex structure involving oligosaccharide chains on the N‐terminal extracellular domain of GPC.