Recombinant antigen diagnostics for filoviruses

DESCRIPTION (provided by applicant): Viral hemorrhagic fevers (VHFs) are serious, often fatal, illnesses characterized by high fever, damage to the vascular system and multiorgan failure. Our team has successfully produced prototype Lassa virus ELISA that are based on recombinant proteins rather than on reagents that must be produced in high containment laboratories. Under the proposed SBIR, we will now perform critical steps in the preclinical development of commercial recombinant antigen diagnostics for filoviruses, including development of point-of-care lateral flow assays. In MILESTONE 1, we will identify recombinant filovirus proteins (Ebola (EBOV) and Marburg (MARV)) GP, sGP (EBOV only), NP and VP40 appropriate for immunoassay development and initiate production of monoclonal antibodies (MAbs) to these proteins. In MILESTONE 2, we will develop prototype recombinant IgM-, IgG-, and antigen-capture ELISA and lateral flow diagnostics for filoviruses (EBOV and MARV). In the projected MILESTONE 3, we will optimize the scale up and purification of both recombinant filovirus proteins and MAbs, as well as convert to manufacturing with Quality Assurance (QA)/Quality Control (QC) to provide quantities of recombinant proteins and MAbs sufficient for development and testing of commercial assays. In MILESTONE 4, we will perform field testing or BSL-4 testing of the recombinant filovirus ELISA and lateral flow assays to validate and compare with PCR assays. MILESTONE 5 will involve converting manufacturing of the assays to Good Manufacturing Practices (GMP) with QA/QC. The potential use of VHF agents such as EBOV and MARV filoviruses (both are BSL-4 and NIAID Category A agents) as biological weapons directed against civilian or military targets necessitates development of effective, highly sensitive and specific, easy to use, adaptable, and cost-effective diagnostics for public health laboratories, hospital-based clinical laboratories, and point-of-care use. The impact of EBOV in Africa is considerable, and effective diagnostics against these and related viruses can also provide a very significant public health benefit. PUBLIC HEALTH RELEVANCE: The potential use of VHF agents such as the filoviruses Ebola (EBOV) and Marburg (MARV; both are BSL-4 and NIAID Category A agents) as biological weapons directed against civilian or military targets necessitates development of effective, highly sensitive and specific, easy to use, adaptable, and cost-effective diagnostics for public health laboratories, hospital-based clinical laboratories, and point-of-care use. Under the proposed SBIR we will perform critical steps in preclinical development of commercial recombinant antigen diagnostics for filoviruses, including development of point-of-care lateral flow assays.