A new method of non-cross-linking conjugation of polysaccharides to proteins via thioether bonds for the preparation of saccharide-protein conjugate vaccines.
Bacterial polysaccharides, including capsular polysaccharides, are poor immunogens particularly in young infants. However, conjugation of bacterial polysaccharides to immunogenic carrier proteins generally results in conjugates that induce strong antipolysaccharide T-helper-cell dependent immune responses, also in young infants. The magnitude of the response and the extent of the T-helper-cell dependency is related to the chemical characteristics of the particular conjugate such as presence or absence of polysaccharide-protein cross-linking, presence or absence of spacer arms, character of spacer arms, type of carrier protein, size of conjugated polysaccharide hapten and molar degree of substitution. In the present study a new, general and simple method for the preparation of poly- and oligosaccharide-protein conjugates is presented. This new method is based on spacer-introducing chemistry that allowed for conjugation of a model polysaccharide, dextran, ranging in size from 0.5 to 150 kDa, to tetanus toxoid (TTd). The developed conjugation method involves derivatization of polysaccharide with 2-iminothiolane (2-IT) and activation of carrier protein, such as TTd, with N-hydroxysuccinimide ester of bromoacetic acid. Reaction rates and accordingly the substitution of the conjugates, could be controlled by varying time, pH and concentration of the reactants. Unlike direct reductive amination, the 2-IT based conjugation technology is fast and made it possible to couple fairly large polysaccharides to TTd.
Pawlowski
A
A
Department of Bacteriology, Swedish Institute for Infectious Disease Control, Stockholm.
Källenius
G
G
Svenson
S B
SB
D003911:Dextrans; D004355:Drug Stability; D006863:Hydrogen-Ion Concentration; D008956:Models, Chemical; D011135:Polysaccharides, Bacterial; D011506:Proteins / Q000138:chemical synthesis*; D013440:Sulfides; D013745:Tetanus Toxoid; D014612:Vaccines / Q000138:chemical synthesis*; D018074:Vaccines, Conjugate
D003911:Dextrans; D011135:Polysaccharides, Bacterial; D011506:Proteins; D013440:Sulfides; D013745:Tetanus Toxoid; D014612:Vaccines; D018074:Vaccines, Conjugate