Novel PhageLock assay to measure hepcidin for clinical monitoring

摘要/摘要 海普西丁是一种负责调节循环铁浓度的多肽激素。过度表达 海普西丁会导致严重的缺铁和贫血，而海普西丁缺乏则会导致铁超载。不 令人惊讶的是，海普西丁的产生和循环浓度的异常与 各种疾病状态，如肾脏疾病、癌症、遗传性血色素沉着症、缺氧、 炎症、感染、炎症性疾病和心脏病。海普西丁的准确检测方法 血清将有助于提高我们对铁代谢的理解、诊断和临床处理。 精神错乱。不幸的是，没有FDA批准的方法来测量血浆中的海普西丁水平。这个 海普西丁的免疫原性很低，阻碍了临床相关海普西丁试验的发展， 这使得制造针对这种荷尔蒙的抗体具有极大的挑战性。为数不多的能做到这一点的免疫分析 EXist在特异性方面的特征很差，他们很难区分庚西丁和多个 在血液循环中发现的荷尔蒙的非活性形式。AffinEnergy计划开发一种基于噬菌体的 临床化学分析将实现对血浆的频繁、简单和负担得起的监测 海普西丁-25水平。使用我们的核心技术噬菌体展示生物扫描，我们已经识别出一种 专有的与海普西丁-25结合的“捕获”肽。在此阶段1应用程序中，我们打算确定一个 并在快速的、 简单易用的夹心酶联免疫吸附试验。在第一阶段结束时，我们希望拥有一种 可在第二阶段放大，供临床医生和研究人员用于测量 血浆。

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.