Novel PhageLock assay to measure hepcidin for clinical monitoring

总结/摘要 铁调素是负责调节循环铁浓度的肽激素。过表达 铁调素导致严重的缺铁和贫血，而铁调素缺乏导致铁超负荷。不 令人惊讶的是，铁调素产生和循环浓度的异常与 各种疾病状态，如肾病、癌症、遗传性血色病、缺氧、贫血、 炎症、感染、炎性疾病和心脏病。铁调素的准确检测方法 血清将导致我们对铁代谢的理解，诊断和临床管理的改善 紊乱不幸的是，没有FDA批准的用于测量血浆中铁调素水平的测定。的 临床相关的铁调素测定法的开发受到铁调素非常低的免疫原性的阻碍， 这使得产生抗这种激素的抗体极具挑战性。为数不多的免疫检测 存在的在特异性方面的特征很差，并且他们很难将hepcidin与多种hepcidin区分开来。 在血液循环中发现的激素的非活性形式。Affinergy计划开发一种基于噬菌体的 临床化学分析，将使频繁，简单，负担得起的监测血浆 铁调素-25水平。利用我们的噬菌体展示生物淘选核心技术， 专有的“捕获”肽结合铁调素-25。在第一阶段的申请中，我们打算确定一个 铁调素-25特异性“检测”噬菌体，并将铁调素-25结合肽和噬菌体快速， 易于使用的基于夹心ELISA的测定。在第一阶段结束时，我们希望有一种技术， 可以在第2阶段扩大规模，供临床医生和研究人员使用，以测量 等离子体

SUMMARY/ABSTRACT Hepcidin is the peptide hormone responsible for regulating circulating iron concentrations. Overexpression of hepcidin results in severe iron deficiency and anemia, whereas hepcidin deficiency leads to iron overload. Not surprisingly, abnormalities in hepcidin production and circulating concentrations are highly correlated with various disease states such as renal disease, cancer, hereditary hemochromatosis, hypoxia, anemia of inflammation, infection, inflammatory diseases, and heart disease. Accurate methods for detecting hepcidin in serum will lead to improvements in our understanding, diagnosis, and clinical management of iron metabolism disorders. Unfortunately, there is no FDA-approved assay for measuring hepcidin levels in plasma. The development of a clinically relevant hepcidin assay has been hindered by hepcidin’s very low immunogenicity, making it extremely challenging to produce antibodies against the hormone. The few immunoassays that do exist are poorly characterized in terms of specificity, and they struggle to differentiate hepcidin from the multiple inactive forms of the hormone that are found in the circulation. Affinergy plans to develop a phage-based clinical chemistry assay that will enable frequent, simple, and affordable monitoring of plasma hepcidin-25 levels. Using our core technology of phage display biopanning, we have already identified a proprietary “capture” peptide that binds to hepcidin-25. In this Phase 1 application, we intend to identify a hepcidin-25 specific “detection” phage and incorporate the hepcidin-25-binding peptide and phage in a rapid, easy-to-use sandwich ELISA-based assay. At the conclusion of Phase 1, we expect to have a technology that can be scaled up in Phase 2 for use by clinicians and researchers to measure hepcidin concentrations in plasma.