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Quantitative Studies of the Specificity of Anti-Pneumococcal Antibodies, Types III and VIII

IV. Binding of Labeled Hexasaccharides Derived from S3 by Anti-S3 Antibodies and Their Fab Fragments¹

From the Biological Laboratories, Harvard University, Cambridge, Massachusetts 02138

Abstract

A method has been described for labeling of oligosaccharides, isolated from partial hydrolysates of Type III pneumococcal polysaccharide, by reduction of the terminal hexose units with tritiated borohydride. Interactions of rabbit anti-S3 antibody and its purified Fab fragments, with purified, labeled and reduced hexasaccharides, (T)3A6 and (T)3D6, derived respectively from S3 by acid and by enzymatic hydrolysis, were studied by equilibrium dialysis. Homogeneous binding was observed in each case with association constants exceeding 10⁵ L/mol. Removal of the Fc fragment followed by reduction and alkylation to 3.6 S fragments did not significantly alter the magnitude of the association constants.

Footnotes

This work was supported by National Science Foundation Grants GB3099 and GB7054.

² Present address: The Hebrew University, Hadassah Medical School, Jerusalem, Israel.

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