Novel antibody based molecules for detection of the microbial pathogen Yersinia pestis
The Gram negative bacterium Yersinia pestis is the aetiological agent of the plague, a disease which throughout history is believed to have caused death and social devastation on a scale unmatched by any other infectious disease. Plague is not an eradicated disease and is still prevalent today. Endemic within animal populations on nearly every continent it causes the death of approximately 2000 humans each year. The recent emergence of multi drug resistant strains along with a potential for illegitimate use as an agent of biological warfare, mean that plague still presents a significant threat to human health. The rapid and sensitive detection of microbial pathogens such as Y. pestis in the environment is essential for the successful implementation of effective countermeasures in order to minimise the outcome of a potential epidemic or biological agent attack. The research described herein investigated the efficacy of recombinant methodologies to generate antibody based molecules against Y. pestis, for employment as reagents in environmental monitoring systems designed to specifically detect and identify airborne pathogens. A total of twelve separate single chain antibody fragments (scAbs) were generated against three individual Y. pestis components, surface exposed or intracellular, either via the rescue and cloning of V genes from existing monoclonals or via the affinity selection of antigen specific antibody fragments from antibody phage display libraries. Of the twelve antibodies generated during this study a high majority (10/12) demonstrated an exceptionally high level of specificity and sensitivity for their particular antigen and initial results from preliminary binding analyses unequivocally established their significance as novel capture reagents within immuno-detection/diagnostic platforms.