Serum Hepcidin Immunoassay: Laboratory to Marketplace

DESCRIPTION provided by applicant Disorders of iron metabolism are among the most common human diseases During the last decade rapid progress has been made in understanding the molecular basis of iron homeostasis and its disorders The peptide hormone hepcidin has emerged as the master regulator of iron metabolism Dysregulation of hepcidin is the principal or contributing factor in most systemic iron disorders In humans primary overproduction of hepcidin due to genetic deficiency of a negative regulator of hepcidin transcription the membrane protein matriptase TMPRSS leads to iron refractory iron deficiency anemia IRIDA IRIDA is a disorder of excessive hepcidin synthesis relative to plasma iron and can be diagnosed by measuring hepcidin levels rather than by DNA sequencing the current confirmatory diagnosis There is a well recognized unmet need for a robust and widely available clinical assay for serum hepcidin but progress towards this goal has been slowed by the poor immunogenicity of hepcidin Intrinsic Life Sciences ILS developed and validated the worldandapos s first RUO serum hepcidin immunoassay competitive ELISA C ELISA that relied on a finite supply of high quality rabbit polyclonal antibodies This first generation hepcidin assay was instrumental in showing that the IRIDA phenotype is caused by inappropriately elevated hepcidin in relation to plasma iron levels and after preliminary data analysis we have demonstrated that this assay can definitively distinguish with high sensitivity and specificity individuals with IRIDA due to TMPRSS mutations from a normal control population of children and adults children and adults with uncomplicated iron deficiency ID and a highly selected population of individuals who have ID that is resistant to oral iron therapy who have been referred to as andquot rule outandquot IRIDA During Phase II research ILS discovered key monoclonal antibodies to hepcidin and developed two robust C ELISAs These prototype new generation andapos wetandapos assays have an excellent dynamic range possess ideal precision and linearity characteristics and display LLOQ LLOD and EC values and intra and inter assay CVandapos s that parallel or exceeds those of our current polyclonal RUO C ELISA Preliminary analysis of de identified IRIDA patient samples have demonstrated that MAb and the NT biotinylated hepcidin tracer yield excellent sensitivity specificity and AUROC that parallel our polyclonal RUO C ELISA We propose to continue to develop the Hepcidin IRidA Competeandquot IVD as a desiccated microplate assay and demonstrate its clinical utility for diagnosis of IRIDA in anemic patients we will enroll Under ISO GMP regulatory guidelines we will complete pre manufacturing Randamp D on our hepcidin IVD finalize design features and produce test lots of assay components confirm reagent performance finalize the Device Master Record and file pre IDE materials to the FDA We will manufacture compliance lots conduct continuous FDA compliant manufacturing performance and quality studies To demonstrate its clinical use we will seek FDA pre IDE guidance finalize design of a prospective clinical trial to be performed at Boston Childrenandapos s Hospital and test the Hepcidin IRidA Competeandquot IVD for diagnostic discrimination between TMPRSS patients with IRIDA and clinically indistinguishable anemic patients enrolled in the trial PUBLIC HEALTH RELEVANCE The founders of Intrinsic Life Sciences ILS pioneered discovery of hepcidin the master iron regulatory hormone in humans and developed the worldandapos s first validated serum hepcidin immunoassay as well as described its key role regulating dietary iron absorption and iron recycling With NIH Phase II funding our team discovered specific monoclonal antibodies MAb to hepcidin and used one MAb to make two prototype serum hepcidin diagnostic tests that are suitable for FDA approval With NIH Phase II B funding ILS will complete pre commercialization Randamp D and select one prototype assay for cGMP ISO manufacturing of the Hepcidin IRidA Competeandquot IVD perform a prospective clinical trial in collaboration with Boston Childrenandapos s Hospital and clinically validate our diagnostic testfor definitive differential diagnosis of a rare genetic iron disorder Iron Refractory Iron Deficiency Anemia IRIDA that has clinical characteristics common in anemic pediatric adolescent patients without the disorder FDA approval of the Intrinsic Hepcidin IRidA Competeandquot IVD will enable rapid diagnosis of IRIDA without expensive genetic testing or extensive clinical work ups