Master of Science (MS)
anti-venom, peptide production
Microbiology; Molecular biology
A cost-effective therapy for snake bites and other biological toxins could save thousands of lives each year. A low-cost therapy can be achieved using a bacterial host that expresses a peptide that prevents death when administered after exposure to these toxins. A protein expressed in opossums, lethal toxin neutralizing factor (LTNF), provides the animal’s resistance to venom and a variety of other toxins. A small peptide fragment of this protein provides the same lifesaving properties as the full protein sequence. A synthetic copy of this peptide was tested in mice exposed to rattlesnake venom and showed lifesaving potential. The peptide was expressed in E. coli and the feasibility of a lower cost process for producing an anti-venom therapy was shown. Two E. coli strains were constructed: snk1 and snk23. This was accomplished by introducing expression vectors containing the peptide sequence alone or a polypeptide sequence that contains 23 repeats of peptide. Once the bacterial cells were transformed with the antivenom DNA, the peptide was produced in shake flasks and scaled up to 2L fermentations. While confirming the expression of the single peptide is ongoing, the larger polypeptide was expressed and recovered in inclusion bodies.
Cifelli, Angela, "Construction of Bacterial Strains Containing a Plasmid Producing an Opossum-Based Anti-Venom Peptide for Low Cost Production of the Peptide" (2017). Master's Theses. 4792.