One-pot preparation of a series of glycoconjugates with predetermined antigen-carrier ratio from oligosaccharides that mimic the O-PS of Vibrio cholerae O:1, serotype Ogawa.
Di-through the pentasaccharide that mimic the upstream terminus of the O-specific polysaccharide of Vibrio cholerae O:1, serotype Ogawa were synthesized in the form of 5-methoxycarbonylpentyl glycosides and linked to BSA using squaric acid diester chemistry. The conjugation reactions were monitored by surface-enhanced laser-desorption/ionization-time-of-flight mass spectrometry (SELDI-TOF MS), which allowed conducting the conjugation of the synthetic oligosaccharides in a controlled way and termination of the reaction when the desired molar hapten-BSA ratio had been reached. This made it possible to prepare, from one hapten in a one-pot reaction, a series of neoglycoconjugates having different, predetermined carbohydrate-carrier ratios. The accuracy of molecular mass determination in SELDI-TOF MS analysis could be increased by using the carrier protein as the internal standard.
Saksena
Rina
R
Laboratory of Medicinal Chemistry, National Institute Diabets and Digestive and Kidney Diseases, National Institutes of Health, Building 8, Rm B1A24, Bethesda, MD 20892-0815, USA.
Ma
Xingquan
X
Kovác
Pavol
P
D000588:Amines / Q000737:chemistry; D000941:Antigens / Q000737:chemistry*; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D006001:Glycoconjugates / Q000138:chemical synthesis* / Q000737:chemistry; D006027:Glycosides / Q000737:chemistry; D006241:Haptens / Q000737:chemistry; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000138:chemical synthesis* / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry; D011135:Polysaccharides, Bacterial / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013997:Time Factors; D014734:Vibrio cholerae / Q000378:metabolism*
D000588:Amines; D000941:Antigens; D006001:Glycoconjugates; D006027:Glycosides; D006241:Haptens; D009844:Oligosaccharides; D011134:Polysaccharides; D011135:Polysaccharides, Bacterial