Production of Recombinant Injectosome and Outer Membrane Proteins from Yersinia Pestis KIM5

摘要

Molecular biology techniques and low cost reagents have lowered the barriers to entry for development of biological arsenals by non-state and state groups. Additionally, genetic engineering of epitope targets from such pathogens as Y. pestis would annul current detection methods and therapeutic treatments. Vaccines often have short shelf lives and are of minimal utility if not used prior to exposure to BW agents. Camelidae, including camels and llamas produce unique antibodies termed Nanobodies(registered) (Nbs) or antigen specific fragments (V(HH)) which are much smaller than traditional antibodies (15 vs. approximately 150 kDal) yet seem to attach with the same selectivity and affinity as full antibodies. V(HH) are more stable than the fragile, more bulky antibodies and maintain their structure and function even at high temperature and humidity. It is thought that, due to these unique characteristics, V(HH) could be reconstituted from a lyophilized pellet and used as a real time injectable immunotherapeutic to be used when warfighters have been exposed to BW. Two logical candidates for V(HH) production are the low-calcium-response V protein (LcrV) and the needle-like Yop (Yersinia Outer- membrane Protein) Secretion Protein F (YscF). This effort successfully produced quantities greater than 1-mg purified native recombinant LcrV and YscF proteins as antigens for V(HH) production.