Synthesis and immunochemical characterization of neoglycoproteins containing epitopes of the inner core region of Pseudomonas aeruginosa RNA group I lipopolysaccharide.
The disaccharide allyl 2-acetamido-2-deoxy-alpha-D-galactopyranosyl-(1-->3)-7-O-carbamoyl-L-glycero-alpha-D-manno-heptopyranoside 5 (GalNAc-cmHep-allyl) was synthesized starting from 1 and 2. Compound 5, cmHep-allyl and the disaccharide cmHep-(1-->3)-Hep-allyl were converted into cysteamine-spacered derivatives and conjugated to bovine serum albumin (BSA) to yield the neoglycoconjugates 7--9, respectively. These conjugates were used to immunize mice and to prepare monoclonal antibodies (mAbs) which were characterized in comparison to mAbs obtained after immunization with heat-killed Pseudomonas aeruginosa strain H4. Two antibodies obtained after immunization with the neoglycoconjugates bound strongly to cmHep-BSA and with lower affinity to cmHep-Hep-BSA but did not bind to GalNAc-cmHep-BSA or to H4 LPS. Another antibody obtained after immunization with heat-killed bacteria bound to LPS and GalNAc-cmHep-BSA but not to cmHep-Hep-BSA or cmHep-BSA
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Institute of Chemistry, University of Agricultural Sciences, Vienna, Austria.
Brade
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Sanchez-Carballo
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Brade
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Kosma
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D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000937:Antigen-Antibody Reactions; D002417:Cattle; D004187:Disaccharides / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology; D004306:Dose-Response Relationship, Immunologic; D004797:Enzyme-Linked Immunosorbent Assay; D006023:Glycoproteins / Q000138:chemical synthesis* / Q000737:chemistry / Q000276:immunology; D007114:Immunization; D008070:Lipopolysaccharides / Q000138:chemical synthesis* / Q000737:chemistry / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D015394:Molecular Structure; D011550:Pseudomonas aeruginosa / Q000737:chemistry*; D012703:Serotyping; D012710:Serum Albumin, Bovine / Q000737:chemistry
D000911:Antibodies, Monoclonal; D004187:Disaccharides; D006023:Glycoproteins; D008070:Lipopolysaccharides; D012710:Serum Albumin, Bovine