This research proposed to advance the technology available for generating and characterizing antibodies specific for carbohydrate structures of biological interest. Towards this goal, we have used recombinant antibody phage display to isolate new monospecific antibody fragments (Fabs) that recognize carbohydrate antigens. We have also attempted to characterize the carbohydrate epitope bound by the antibodies and to take advantage of the recombinant system to make these Fabs, and other forms of recombinant antibodies, more useful. Research conducted this year revealed problems associated with light-chain expression and assembly in the pComb3 system we have been using to generate recombinant Fabs. We have overcome this problem by switching to the single-chain Fv expression system (pCANTAB6) of Cambridge Antibody Technology. Using this technology, we have begun a study of how glycosylation of human antibodies plays a role in antigen binding and the relationship of these effects on human diseases such as primary amyloidosis and rheumatoid arthritis.
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