1a.Objectives (from AD-416):
The goal of this collaborative research project is to produce camel monoclonal antibodies (cMAb) against Foot-and-Mouth Disease Virus (FMDV) non-structural proteins (NSP) and virus particles (SP). USDA, ARS and Ben Gurion University scientists will design a reagent for viruses that will target three relevant FMDV serotyes including O, A and the South African Territories (SAT) types of Foot-and-Mouth Disease Virus (FMDV). An additional objective includes an attempt to assemble diagnostics assays using selected virus-specific cMAb developed in this project. Currently, there are no effective “in house” and affordable surveillance diagnostic test widely available in sub-Saharan Africa.
1b.Approach (from AD-416):
Virus specific camel monoclonal antibodies (cMAbs) in camelids directed against the Foot-and-Mouth Disease Virus (FMDV) structural proteins (SP) and non-structural proteins (NSPs) will be produced. Using an eukaryotic expression system tailored to the FMDV proteins, reactive hybridoma clones to the capsid, the 3D and 3ABC proteins will be screened and selected. Additionally, efforts will be aimed at the development of suitable and easy to deploy DIVA (differentiation between infected and vaccinated animals) (DIVA) tests utilizing the camel MAbs in a test platform. The formulated test/s will be bench standardized to assess the rapid and accurate detection of anti-NSP and FMDV detection. The specificity, stability and affinity characteristics of camel MAbs make them excellent choices in biosensor applications and they will potentially be more stable reagents when applied in a validated test.
During FY 2013, camels were maintained and we expressed sufficient soluble recombinant viral protein 1 (VP1) in bacteria from FMD strains O and SAT 1 and optimized the camel vaccination protocol. In final analysis, it was determined that the following protocol was optimal: one injection of 1 mg/injection in Freund’s complete adjuvant followed by eight weekly injections with 1 mg/injection in Freund’s incomplete adjuvant. These results demonstrated that serum titers increased during the course of the protocol and at the termination, the serum titers of specific antibodies to VP1 were sufficiently high for generation of recombinant phage displayed camel monoclonal antibody libraries.
No technologies were transferred in FY 2013. No publications have been produced during this fiscal year.