title: Neoglycoproteins as carbohydrate antigens: synthesis, analysis, and polyclonal antibody response.
abstract: The analysis and polyclonal antibody response for newly synthesized maltose-BSA conjugate neoglycoproteins is described. In this first proof of concept study, a simple carbohydrate antigen, maltose, was linked to BSA by reductive amination. An aglycone spacer was utilized to conserve the intact annular maltose structure and to promote the accessibility of the carbohydrate immunogen hapten during immunization. The neoglycoproteins were investigated by CGE and the number of conjugated maltose residues was determined by MALDI-TOF MS. The neoglycoproteins were then evaluated by immunization of BALB/c mice and the polyclonal antibody response was tested by ELISA as evidence for the presence of sugar-containing epitope-specific antibodies. Selective antibody binding was demonstrated to the synthesized neoglycoproteins with different (low and high) glycosylation degrees suggesting the possible use of this approach to generate antibodies. Moreover, the polyclonal antibody response was not inhibited by maltose or other simple carbohydrates to confirm presence of the neoglycoprotein-specific antibodies.
authors.affiliation: Horváth Laboratory of Bioseparation Sciences, University of Debrecen, Debrecen, Hungary.
authors.forename: Márta
authors.initials: M
authors.lastname: Kerékgyártó
authors.forename: Anikó
authors.initials: A
authors.lastname: Fekete
authors.forename: Zoltán
authors.initials: Z
authors.lastname: Szurmai
authors.forename: János
authors.initials: J
authors.lastname: Kerékgyártó
authors.forename: László
authors.initials: L
authors.lastname: Takács
authors.forename: István
authors.initials: I
authors.lastname: Kurucz
authors.forename: András
authors.initials: A
authors.lastname: Guttman
mesh_terms: D000818:Animals; D000906:Antibodies / Q000097:blood*; D000941:Antigens / Q000032:analysis* / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D002417:Cattle; D019075:Electrophoresis, Capillary; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D006023:Glycoproteins / Q000032:analysis / Q000737:chemistry* / Q000276:immunology* / Q000378:metabolism; D008320:Maltose / Q000737:chemistry / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D000906:Antibodies; D000941:Antigens; D000939:Epitopes; D006023:Glycoproteins; D012710:Serum Albumin, Bovine; D008320:Maltose
keywords: Carbohydrate antigen; Carbohydrate-specific antibody; ELISA; Neoglyco-protein