Isolation, characterization and implications of anti-TF (Thomsen-Friedenreich) agglutinins from different sources.
Anti-TF agglutinins from peanut (Arachis hypogaea) and from vertebrate sera of different species have been successfully isolated by affinity chromatography on acid-activated Sepharose 4 B. The proteins were characterized by immunoelectrophoresis, polyacrylamide gel electrophoresis in the presence of SDS and with respect to their carbohydrate binding specificities. Anti-TF substances from sera showed one precipitin arc in immunoelectrophoresis, but quantitative immunoprecipitation revealed our human anti-TF to be a mixture of the three Ig-classes IgG, IgA and IgM. This finding was confirmed on SDS gel electrophoresis, where high molecular weight aggregates were found before reduction. Hemagglutination inhibition revealed that all isolated anti-TF compounds exhibit an exceptionally high affinity for the immunodominant group of the TF-antigen, namely the beta-D-galactosyl-(1 leads to 3)-N-acetyl-D-galactosamine disaccharide. On examination of formalin-fixed and neuraminidase treated tissue sections (kidney, mammary gland), fluorescein-labelled anti-TF from horse serum showed a virtually identical pattern when compared with fluorescein labelled peanut lectin. Likewise isolated IgA-class myeloma J 539, which shows specificity against beta-(1 leads to 6)-galactans, only bound to the appropriate Gal-beta-(1 leads to 6)-Gal structures, such as those found on bovine lung or the albumin gland of Helix pomatia. Rabbit anti-VCN (Vibrio cholerae neuraminidase) activity could be selectively abolished by beta-galactosyl-containing inhibitors, whereas papain F(ab) fragments from rabbit anti-VCN immunoglobulin did not compete with anti-TF for binding sites on VCN-treated human red cells. Anti-TF, on the other hand, did not compete with anti-VCN for active VCN.
D000373:Agglutinins / Q000302:isolation & purification*; D000818:Animals; D000918:Antibody Specificity; D010367:Arachis; D004187:Disaccharides / Q000276:immunology; D006385:Hemagglutination Inhibition Tests; D006736:Horses; D006801:Humans; D007106:Immune Sera / Q000494:pharmacology; D008951:MNSs Blood-Group System; D051379:Mice; D009194:Myeloma Proteins / Q000276:immunology; D000509:alpha-Fetoproteins
D000373:Agglutinins; D004187:Disaccharides; D007106:Immune Sera; D008951:MNSs Blood-Group System; D009194:Myeloma Proteins; D000509:alpha-Fetoproteins