Generation of systems for the synthesis of recombinant proteins for use in rapid

Lead Research Organisation: University of Kent


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Studentship Projects

Project Reference Relationship Related To Start End Student Name
BB/M017060/1 30/09/2015 29/09/2019
1666804 Studentship BB/M017060/1 30/09/2015 29/09/2019
Description We have achieved expression and purification of soluble Elastase, a secreted protease and known virulence factor of the gram negative bacterium Pseudomonas aeruginosa.

We have successfully generated sheep and rabbit polyclonal antibodies to Elastase. These antibodies have been incorporated into two diagnostic assays; a lateral flow assay and an enzyme-linked immunosorbent assay, able to detect Elastase at concentrations of < 100 pg/mL with no observed cross-reactivity with other pseudomonas proteins or proteins secreted by other gram negative bacteria. This assay has been tested on clinical isolates obtained from cystic fibrosis patients in collaboration with Imperial College London.

We have attempted to generate high affinity monoclonal Fab and scFv fragments specific to elastase via antibody phage display. However, despite generating large, diverse immune libraries, we have currently been unable to successfully select soluble monoclonal fragments.
Exploitation Route The anti-elastase diagnostic assay that we have been developing, once finished, would ideally be used in the clinic to detect early stage Pseudomonas aeruginosa infections in Cystic Fibrosis patients. However, more work must be performed using clinical samples in order to determine if it is a viable assay.

Further development of the assay could be performed through the generation of monoclonal antibody fragments via antibody phage display, as well as generation of antibodies to other pseudomonas virulence factors in order to produce a multiplex assay, if the detection of elastase alone is not considered a reliable diagnostic test.
Sectors Healthcare,Pharmaceuticals and Medical Biotechnology