Conjugation of a peptide to mannan and its confirmation by tricine sodium dodecyl sulfate–polyacrylamide gel electrophoresis

The conjugation of polysaccharides to peptides is essential for antigen delivery and vaccine development. Herein, we show that tricine SDS-PAGE in combination with Coomassie Blue staining was adequate to determine the conjugation efficacy of a peptide (epitope 35–55 of myelin oligodendrocyte glycopr...

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Bibliographic Details
Published in:Analytical biochemistry 2015-09, Vol.485, p.43-45
Main Authors: Tapeinou, Anthi, Androutsou, Maria-Eleni, Kyrtata, Konstantina, Vlamis-Gardikas, Alexios, Apostolopoulos, Vasso, Matsoukas, John, Tselios, Theodore
Format: Article
Language:English
Subjects:
Conjugation
Electrophoresis, Polyacrylamide Gel - methods
Glycine - analogs & derivatives
Glycine - chemistry
Mannan
Mannans - chemistry
MOG35–55
Myelin oligodendrocyte glycoprotein
Myelin-Oligodendrocyte Glycoprotein - chemistry
Peptide
Peptide Fragments - chemistry
Tricine SDS-PAGE
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Summary:The conjugation of polysaccharides to peptides is essential for antigen delivery and vaccine development. Herein, we show that tricine SDS-PAGE in combination with Coomassie Blue staining was adequate to determine the conjugation efficacy of a peptide (epitope 35–55 of myelin oligodendrocyte glycoprotein) to mannan. In addition, tricine SDS-PAGE and periodic acid–Schiff stains were able to monitor the redox state of mannan. Using the described protocol, more than 99.9% of a peptide containing five lysines at its N-terminus was confirmed conjugated to mannan.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2015.06.010