Reagents to Enhance Detection of Raw and Biologically Transformed Nanomaterials

描述(由申请人提供)：在工业和消费应用中越来越多地使用工程纳米材料，这引起了人们对其环境健康安全的严重关注(EH和S)。一个特别令人担忧的问题是，职业暴露于高度分散的球形和纤维状的原材料。研究人员试图量化纳米材料与生物系统的相互作用，并开发纳米颗粒EH和S风险的预测模型。然而，所使用的模型系统施加的限制、NP标准的缺乏、研究人员未能量化或报告所研究NP的物理化学性质、以及灵敏的分析仪器的不足，共同阻碍了实现EH和S目标的进展。在拟议的项目中，我们试图通过开发一套试剂(NProbe)来克服检测纳米颗粒的局限性，该试剂将增强纳米颗粒在生物组织中的存在，这些生物组织由于其大小可能被其他方法掩盖。NProbe试剂将以类似于标准免疫组织学分析中用于检测和定量蛋白质的方式发挥作用。利用基于蛋白质的(抗体和纤维连接蛋白)噬菌体展示技术，将产生与原始NP和转化的纳米材料都具有高结合亲和力的NProbe。感兴趣的纳米材料包括碳纳米管、金属氧化物、金属和半导体。我们将验证NProbe与原始纳米材料的结合，并测试它们与相关和不同组成的样品的交叉反应性。N还将展示通过渗透皮肤转变的纳米材料与nProbe的结合。我们相信，nProbe试剂将有助于研究人员评估纳米颗粒在所有哺乳动物、脊椎动物和无脊椎动物系统中的组织相互作用，以及检测环境中的纳米颗粒。NProbe试剂的成功开发将对纳米颗粒风险评估研究产生革命性的影响。 与公共健康相关：在过去的几年里，工程纳米材料在工业和消费应用中的使用速度加快，引发了人们对纳米颗粒暴露带来的环境健康安全风险的严重担忧(EH&S)。敏感的分析仪器供应不足造成的限制，造成了我们在量化工程纳米材料的命运和运输方面的能力差距。在这个拟议的项目中，我们通过开发一套试剂(NProbe)来解决这个问题，该试剂将能够检测纳米颗粒、皮肤和其他生物或环境系统中的纳米颗粒。NProbe试剂将以类似于标准免疫组织学分析中用于检测和定量蛋白质的方式发挥作用。利用基于蛋白质的(抗体和纤维连接蛋白)噬菌体展示技术，将产生与原始NP和转化的纳米材料都具有高结合亲和力的NProbe。

DESCRIPTION (provided by applicant): The increasing use of engineered nanomaterials in industrial and consumer applications has raised serious concerns about their environment health safety (EH&S). A particular concern is for occupational exposure to raw materials in the form of highly dispersed spherical and fiber- like shapes. Researchers seek to quantify nanomaterial interactions with biological systems and to develop predictive models of nanoparticle EH&S risk. However, limitations imposed by the model systems employed, the lack of NP standards, the failure of researchers to quantify or report the physiochemical properties of the NP investigated, and the inadequate availability of sensitive analytical instrumentation, have collectively hampered progress in attaining these EH&S goals. In the proposed project we seek to overcome limitations in detecting nanoparticles by developing a set of reagents (NProbes) that will enhance their presence in biological tissues that due to their size may be obscured from detection by other means. NProbe reagents will function in a similar fashion to those used to detect and quantify proteins in standard immunohistological analysis. NProbes with high binding affinity to both raw NP and transformed nanomaterials will be generated using protein-based (antibody and fibronectin) phage display technology. Nanomaterials of interest include carbon nanotubes, metal oxides, metals, and semiconductors. We will validate the binding of NProbes to raw nanomaterials and test their cross reactivity to samples of related and dissimilar composition. NProbe binding to nanomaterials transformed by penetration through skin will be also be demonstrated. We believe that NProbe reagents will be useful to researchers assessing nanoparticles tissue interactions in all mammalian, vertebrate, invertebrates systems as well as detection of nanoparticles in the environment. Successfully development of NProbe reagents will have a transformative effect on nanoparticle risk assessment research. PUBLIC HEALTH RELEVANCE: Over the past few years the use of engineered nanomaterials in industrial and consumer applications has accelerated, raising serious concerns about the environment health safety (EH&S) risk from nanoparticle exposure. Limitations imposed by inadequate availability of sensitive analytical instrumentation have contributed to gaps in our ability to quantify the fate and transport of engineered nanomaterials. In this proposed project we address this issue by developing a set of reagents (NProbes) that will enable detection of nanoparticles skin and in other biological or environmental systems. NProbe reagents will function in a similar fashion to those used to detect and quantify proteins in standard immunohistological analysis. NProbes with high binding affinity to both raw NP and transformed nanomaterials will be generated using protein-based (antibody and fibronectin) phage display technology.