title: A novel synthesis of alpha-D-Galp-(1-->3)-beta-D-Galp-1-O-(CH2)3-NH2, its linkage to activated matrices and absorption of anti-alphaGal xenoantibodies by affinity columns.
abstract: Pig organs transplanted into primates are rapidly rejected because of the interaction between Gal alpha(1-->3)Gal epitopes carried by the graft and natural antibodies (anti-alphaGal antibodies) present in the blood of the recipient. This report describes a simplified synthesis of the xenogeneic disaccharide and its linkage to activated gel matrices. The digalactosides alpha-D-Galp-(1-->3)-alpha,beta-D-Galp-OAll were synthesized by the condensation of the trichloroacetimidoyl 2,3,4,6-tetra-O-benzyl-beta-D-galactopyranoside donor with the 3,4-unprotected allyl 2,6-di-O-benzyl-alpha- or beta-D-galactopyranoside acceptor precursor. Deallylation and hydrogenolysis led to the free digalactoside, whereas hydrogenolysis alone resulted in the 1-O-propyl digalactoside. Both products were tested by inhibition ELISA of natural anti-Gal alpha(1-->3)Gal antibodies. The alpha-D-Galp-(1-->3)-beta-D-Galp-OPr was found to be the best inhibitor. Thus, the allyl group of the partially benzylated alpha-D-Galp-(1-->3)-beta-D-Galp-OAll was engineered, via the hydroxy-, the tosyloxy- and the azidopropyl intermediates, into an aminopropyl group amenable to binding to N-hydroxysuccinimide-activated agarose gel matrices in order to obtain specific immunoabsorption columns. Columns made of gel substituted with 5 micromol of disaccharide per milliliter were found efficient for the immunoabsorption of anti-alphaGal antibodies from human plasma.
authors.affiliation: Unité de Recherche sur l'Immunointervention dans les allo- et les xénotransplantations, INSERM Unité 437, Centre Hospitalier Universitaire de Nantes, France.
authors.forename: J
authors.initials: J
authors.lastname: Liaigre
authors.forename: D
authors.initials: D
authors.lastname: Dubreuil
authors.forename: J P
authors.initials: JP
authors.lastname: Pradère
authors.forename: J F
authors.initials: JF
authors.lastname: Bouhours
mesh_terms: D002240:Carbohydrate Sequence; D002846:Chromatography, Affinity; D004187:Disaccharides / Q000138:chemical synthesis*; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D005690:Galactose / Q000097:blood / Q000737:chemistry / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry; D007075:Immunoglobulin M / Q000737:chemistry; D007163:Immunosorbent Techniques; D008956:Models, Chemical; D008969:Molecular Sequence Data; D011437:Propylamines / Q000138:chemical synthesis*; D012685:Sepharose / Q000737:chemistry; D013388:Succinimides / Q000737:chemistry; D014183:Transplantation, Heterologous
chemical_list: C415344:3-aminopropyl galactopyranosyl-1-3-galactopyranoside; D004187:Disaccharides; D000939:Epitopes; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D011437:Propylamines; D013388:Succinimides; D012685:Sepharose; C001426:N-hydroxysuccinimide; D005690:Galactose