DIRECTING ANTIGEN-SPECIFICITY TOWARDS BOTULINUM NEUROTOXIN WITH COMBINATORIAL PHAGE DISPLAY LIBRARIES

The production of antibodies towards antigens with low immunogenicity is enhanced by the intrinsic efficiency of screening combinatorial lib raries of immunoglobulins. The need to isolate clones with rare bindin g specificities has dictated a highly efficient method of screening an d isolating antibody clones. The production of recombinant immunoglobu lin libraries in bacteria allows for a more controlled selection of an tibody specificity and can be used in circumstances where hybridoma fu sions are unable to isolate rare clones with the desired epitope speci ficity. Botulinum neurotoxin (NT) with associated non neurotoxin prote ins (non-NT) as a complex was used to immunize mice to obtain mRNA for the production of a recombinant antibody library with a repertoire of specificities. Initial screens of the combinatorial library revealed clones which recognized the non-neurotoxin proteins of the toxin compl ex similar to monoclonal antibodies produced by conventional hybridoma fusions. The combinatorial library was re-screened in order to isolat e antibodies that specifically recognized the neurotoxin component of the toxin complex. The ability to alter the biopanning selection proce ss affords the researcher a measure of control in the selection proces s not available with traditional hybridoma fusions.