Investigation of tetanus toxin fragment C for bioconjugation studies with self-assembling vesicle polymers and stimuli responsive materials.

Dominic J. Rucco, Amanda M. Pritzlaff, Daniel A. Savin

Department of Chemistry, University of Florida

In this work we seek to exploit the targeting and cell entry mechanisms of tetanus toxin fragment C by polymer conjugation to self-assembled vesicles in an effort to improve local drug delivery in cancer therapy.  Tetanus toxin fragment C (H_c) provides an example whereby complex and specific actions are achieved through well-orchestrated mechanisms taking advantage of a distinguished protein domain.  In short, recognition of cell bound gangliosides, specifically trisialoganglioside GT1b, triggers receptor mediated endocytosis and subsequent uptake of the protein.  By utilizing the targeting domain of tetanus toxin, this process can be achieved without the toxicity of the native protein. Currently, we are developing protocols for bioconjucation and ultimate study of H_c-polymer systems, and herein we present the overexpression and manipulation of the neuronal targeting domain H_c from E. coli for eventual polymer bioconjugation.