The interaction of mouse myeloma immunoglobulin S15 with negatively charged polysaccharide antigens.
A murine BALB/c IgG2a (lambda 3) myeloma immunoglobulin SAPC-15 with binding activity for negatively charged polysaccharides has been purified by affinity chromatography, and its interaction with heparin and various other polyanionic antigens has been studied. The antigen-binding activity has been demonstrated to reside in the Fab part of the immunoglobulin. The S15 myeloma protein in 0.05 M Tris buffer at pH 7.4 precipitated dextran sulfate, heparin, chondroitin sulfate A, B and C, hyaluronic acid, H. influenzae type b polysaccharide, calf thymus DNA, Klebsiella polysaccharide K63 and poly-L glutamic acid. Of these antigens only dextran sulfate was precipitated in 0.01 M phosphate buffered saline (0.15M), pH 7.4. The pepsin S15 Fab fragment did not precipitate with any of these antigens. The intrinsic tryptophanyl fluorescence of S15 was changed maximally by the addition of heparin, and the binding affinity of the immunoglobulin for this antigen was high (greater than 10(6) L/M). S15 may resemble antibody molecules that react with antigens under non-physiological conditions or in pathological conditions or in the external environment as in the lumen of the gut. All the above interactions of S15 with antigens persisted in 0.05 M Tris buffer made physiologically isotonic by the addition of sucrose, and S15 could thus be used to identify these antigens on cell surfaces.
D000818:Animals; D000941:Antigens / Q000276:immunology; D002850:Chromatography, Gel; D006493:Heparin / Q000276:immunology; D005779:Immunodiffusion; D007122:Immunoelectrophoresis; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D007136:Immunoglobulins / Q000276:immunology* / Q000302:isolation & purification; D051379:Mice; D008807:Mice, Inbred BALB C; D009194:Myeloma Proteins / Q000276:immunology* / Q000302:isolation & purification; D009994:Osmolar Concentration; D010206:Papain; D011135:Polysaccharides, Bacterial / Q000276:immunology*
D000941:Antigens; D007140:Immunoglobulin Fab Fragments; D007136:Immunoglobulins; D009194:Myeloma Proteins; D011135:Polysaccharides, Bacterial; C017551:myeloma immunoglobulins; D006493:Heparin; D010206:Papain