Application of cystamine and N,N'-Bis(glycyl)cystamine as linkers in polysaccharide-protein conjugation.
Pneumococcal polysaccharide type 6B, 14, or 23F (35-70 kDa) was activated with cyanogen bromide and modified with cystamine. After reduction of the spacer, the thiol-containing (i.e. cysteamine-modified) polysaccharide obtained was added in a 5-10-fold molar excess to bromoacetylated tetanus toxoid to give thioether-linked polysaccharide-protein conjugates in a yield of 10-20%. This approach failed for preparing a type 19F polysaccharide-protein conjugate, possibly due to intramolecular elimination of cysteamine from the reduced 19F polysaccharide. When N,N'-bis(glycyl)cystamine was introduced as a spacer molecule, the elimination of the reduced spacer was suppressed, thus allowing preparation of a 19F polysaccharide-tetanus toxoid conjugate (15%).
de Weers
O
O
National Institute of Public Health and the Environment (RIVM), Laboratory of Vaccine Development and Immune Mechanisms, The Netherlands.
Beurret
M
M
van Buren
L
L
Oomen
L A
LA
Poolman
J T
JT
Hoogerhout
P
P
D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D003488:Cyanogen Bromide / Q000378:metabolism; D003538:Cystamine / Q000031:analogs & derivatives* / Q000138:chemical synthesis / Q000737:chemistry*; D005998:Glycine / Q000031:analogs & derivatives; D006001:Glycoconjugates / Q000138:chemical synthesis* / Q000276:immunology; D008969:Molecular Sequence Data; D015394:Molecular Structure; D011135:Polysaccharides, Bacterial / Q000276:immunology / Q000378:metabolism; D013296:Streptococcus pneumoniae / Q000276:immunology*; D013745:Tetanus Toxoid / Q000737:chemistry / Q000276:immunology
D006001:Glycoconjugates; C112701:N,N'-bis(glycyl)cystamine; D011135:Polysaccharides, Bacterial; D013745:Tetanus Toxoid; D003488:Cyanogen Bromide; D003538:Cystamine; D005998:Glycine