Neoglycoproteins as carbohydrate antigens: Synthesis, analysis, and polyclonal antibody response

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 annu...

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Bibliographic Details
Published in:Electrophoresis 2013-08, Vol.34 (16), p.2379-2386
Main Authors: Kerékgyártó, Márta, Fekete, Anikó, Szurmai, Zoltán, Kerékgyártó, János, Takács, László, Kurucz, István, Guttman, András
Format: Article
Language:English
Subjects:
Accessibility
Animals
Antibodies
Antibodies - blood
Antigens
Antigens - analysis
Antigens - chemistry
Antigens - immunology
Antigens - metabolism
Carbohydrate antigen
Carbohydrate-specific antibody
Carbohydrates
Cattle
Conjugates
Electrophoresis, Capillary
ELISA
Enzyme-Linked Immunosorbent Assay
Epitopes
Glycoproteins - analysis
Glycoproteins - chemistry
Glycoproteins - immunology
Glycoproteins - metabolism
Immunization
Maltose
Maltose - chemistry
Maltose - metabolism
Mice
Mice, Inbred BALB C
Neoglyco-protein
Serum Albumin, Bovine - chemistry
Serum Albumin, Bovine - immunology
Serum Albumin, Bovine - metabolism
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
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Summary: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.
ISSN:0173-0835
1522-2683
DOI:10.1002/elps.201300052