1a.Objectives (from AD-416)
We will produce a population of scFv molecules with activity against Ca. Liberibacter asiaticus strains found in Florida as well as other species. We will also produce monoclonal antibodies with activity against the same organisms.
1b.Approach (from AD-416)
Liberibacter will be obtained from infected citrus psyllids (or culture if available). Extracts will be prepared (Flordia strains first) and Liberibacter quantified by Q-PCR to provide a known inoculum to mice. Mice will be sacrificed and scFv antibody library in phage will be prepared following standard methods. The phage library will be enriched for Liberibacter specific scFv by several cycles of removing phage that bind healthy psyllid extracts followed by amplification of those that do not. Monoclonal antibody expressing cell lines will be prepared using hybridoma technology in parallel with the scFv work. The antibodies against Liberibacter will then be used to develop research and diagnostic tests needed to manage the disease beginning with ELISA and dot blot tests and continuing with tests in the 'dip stick' format.
We have been able to isolate and characterize more than 20 unique antibody molecules. All are directed at different defined proteins from ‘Ca. Liberibacter asiaticus’, the pathogen associated with citrus greening HLB in Florida. These antibody molecules are immediately useful for research purposes and will be tested in several novel diagnostic applications.