Affinity chromatography of human anti-dextran antibodies. Isolation of two distinct populations.
Affinity chromatography is a very efficient method for antibody purification. Two affinity chromatography supports were prepared to analyze the specificity of anti-dextran antibodies. Silica beads were grafted with native dextran or with functionalized dextran. The anti-dextran antibodies present in some human sera were analyzed by enzyme-linked immunosorbent assay method. These antibodies play an important role in severe dextran-induced anaphylactic reactions in humans by forming immune complexes with clinical dextran. The results indicated that two distinct populations of anti-dextran antibodies were purified from human serum, using dextran-coated silica beads. Elution from this support with an oligo-dextran of 4000 g/mol allowed the isolation of one population that only recognized native dextran as antigen. Functionalized dextran coated on dextran silica beads led to the purification, with a glycine-HCl buffer, of another subclass of antibodies that recognized substituted dextran derivatives. Furthermore, these antibodies could be useful tools for in vitro and in vivo investigations using dextran derivatives as bio-active polysaccharides.
Anastase
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Laboratoire de Recherches sur les Macromolécules, CNRS URA 502, Institut Galilée, Université Paris Nord, Villetaneuse, France.
Letourneur
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D
Jozefonvicz
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J
D000906:Antibodies / Q000097:blood / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods*; D003911:Dextrans / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D012822:Silicon Dioxide
D000906:Antibodies; D003911:Dextrans; D012822:Silicon Dioxide