The research will result in new kits for quantitative detection and monitoring of hypoxia inducible factor-1 alpha (Hif-1a), a promising cellular target protein for pharmacological intervention. During the Phase 1 project, monoclonal antibodies will be developed and used to create highly sensitive ELISA assays for Hif-1a. Mice immunized with peptides derived from Hif-1a will be used to produce hybridomas secreting anti-Hif antibodies. The prototype assay will be developed in Phase I and characterized using model systems of synthetic/recombinant targets and cultured cells, and SOPs will be created for production of all reagents and materials and for carrying out the assay. The prototype assay will be tested and reduced to practice using biological samples during Phase II. The ELISA assays that will be developed will be able to distinguish between the full-length active form of Hif-1a and the truncated form thought to act as a dominant negative regulator. The proposed ELISA assays will be used to evaluate candidate chemotherapeutic agents thought to affect Hif-1a. The successful development of the proposed assays will help to achieve the goals of the NCI Strategic Plan to support the more efficient evaluation and identification of targeted anti-tumor drugs.
