BIOMONAR: Biosensor nanoarrays for environmental monitoring

BIOMONAR：用于环境监测的生物传感器纳米阵列

• 批准号: 1064267
• 负责人: Antje Baeumner
• 金额: $ 18万
• 依托单位: Cornell University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2015-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1064267&HistoricalAwards=false
• 关键词: BIOMONAR; Biosensor; nanoarrays; environmental; monitoring

1064267BaeumnerEnvironmental samples are complex and it is unrealistic to aim for a "one size fits all" biosensor for the detection of environmental analytes. At the same time, a non-systematic approach to biosensor design, resulting in a suite of very diverse sensors with few common features, hinders the necessary conceptual and fundamental understanding of environmental processes and the design of sensors for new targets. Thus, a group of European researchers and PI Baeumner have been funded by the European Commission in 2010 (for 4 years) for the development of a battery of nanoarray biosensors for the measurement of a spectrum of chemical and biological parameters. Development and fundamental studies of novel biorecognition elements, investigation of reactivities and fluxes of target analytes provide the fundamental understanding needed for the design of sensors for new analytes in complex environmental samples. Specifically, focus is on the process of developing three complementary sensing platforms (optical, liposomal, whole cell) integrating a single family of sensing proteins as biorecognition element capable of sensing an unlimited number of pollutants. Bacterial periplasmic binding proteins (PBPs) are genetically engineered to bind specifically and sensitively to a wide range of analytes relevant to environmental analysis including heavy metals, pesticides and pathogens.The following studies are proposed here in order to gain fundamental understanding of a liposome-based PBP sensing platform prior to its application via the EU project: (1) fundamental understanding of PBPs as biorecognition elements using surface plasmon resonance (SPR), including affinity and kinetic binding studies, (2) development of liposome-based PBP microtiter plate fluorescence assays gaining information on PBP as biorecognition element in effective bioassays in environmental matrices with a quantified characterization of the proteins, (3) development of electrochemiluminescence-based microfluidic liposome biosensors for sensitive environmental analysis avoiding matrix-related non-specific signals. This complements studies carried out with EU partners including (4) surface characterization of PBP-bound liposomes using atomic force microscopy (AFM) and determination of the PBP-analyte binding strength, (5) studies to determine the ability of liposomes to enhance Mach-Zehnder interferometer sensitivity, and (6) the investigation of liposomes as cell-mimics in hydrogels assisting in the study of bioavailability of inorganic and organic compounds.The scientific merit of the specific studies proposed here is the gaining of fundamental knowledge about novel biorecognition approaches leveraging genetically engineered PBPs developed by EU partners. Also, highly sensitive and discriminative environmental biosensors will be developed through a liposome-based electrochemiluminescence (ECL) microfluidic strategy. In addition, via intensive interactions with the EU partners, liposomes will be investigated as multifunctional particles. The scientific merit of the overall studies by the EU partners and PI Baeumner is based on the multidisciplinary approach to developing a generic dynamic framework for quantitative interpretation of the "exposure to effect" chain of processes that determine the biological impacts of pollutants.The broader impact includes (1) the construction and testing of a new generation of biosensors for environmental monitoring by commercial environmental monitoring agency partners within the EU team. This provides direct opportunities for commercialization and transfer of knowledge to relevant end users of the produced technology. Data will be provided directly to the EU and may help in the setting of environmental policies. (2) Organization of workshops for end-users, and intensive training courses for graduate students and postdocs. (3) In addition, PI Baeumner will ensure training of undergraduate students in biosensing research and continue her outreach to high school students with an Indian Tribal High Schools in the upstate NY area.

1064267 Baeumner环境样品是复杂的，并且旨在“一刀切”的生物传感器用于检测环境分析物是不现实的。与此同时，一个非系统的方法来设计生物传感器，导致一套非常不同的传感器，很少有共同的特点，阻碍了必要的概念和基本的理解环境过程和传感器的设计新的目标。因此，一组欧洲研究人员和PI Baeumner在2010年获得了欧盟委员会的资助（为期4年），用于开发一系列纳米阵列生物传感器，用于测量化学和生物参数的光谱。新型生物识别元件的开发和基础研究，目标分析物的反应性和通量的调查提供了复杂环境样品中新分析物的传感器设计所需的基本理解。具体而言，重点是开发三个互补的传感平台（光学，脂质体，全细胞）的过程中，集成了一个单一的家庭的传感蛋白作为生物识别元件能够检测无限数量的污染物。细菌周质结合蛋白（PBP）是一种基因工程蛋白，可以特异性和敏感性地结合多种与环境分析相关的分析物，包括重金属、农药和病原体。为了在欧盟项目中应用基于脂质体的PBP传感平台之前获得基本的了解，本文提出了以下研究：（1）使用表面等离子体共振（SPR）对PBPs作为生物识别元件的基本理解，包括亲和力和动力学结合研究，（2）脂质体的研制基于PBP的微量滴定板荧光测定获得关于PBP作为环境基质中的有效生物测定中的生物识别元件的信息，具有蛋白质的定量表征，（3）开发基于电致发光的微流控脂质体生物传感器，用于灵敏的环境分析，避免了与基质相关的非特异性信号。 这补充了与欧盟合作伙伴开展的研究，包括（4）使用原子力显微镜（AFM）对PBP结合的脂质体进行表面表征并测定PBP-分析物结合强度，（5）测定脂质体增强Mach-Zehnder干涉仪灵敏度的能力的研究，（6）脂质体作为细胞-水凝胶中的模拟物有助于研究无机和有机化合物的生物利用度。这里提出的具体研究的科学价值是获得关于利用欧盟合作伙伴开发的基因工程PBPs的新型生物识别方法的基本知识。此外，高灵敏度和区分环境生物传感器将通过基于脂质体的电化学发光（ECL）微流控策略开发。此外，通过与欧盟合作伙伴的密切相互作用，脂质体将作为多功能颗粒进行研究。欧盟合作伙伴和PI Baeumner的总体研究的科学价值是基于多学科方法，以制定一个通用动态框架，用于定量解释确定污染物生物影响的“暴露于影响”过程链。欧盟团队内的商业环境监测机构合作伙伴构建和测试新一代环境监测生物传感器。这为商业化和向所生产技术的相关最终用户转让知识提供了直接机会。数据将直接提供给欧盟，并可能有助于制定环境政策。(2)为最终用户举办讲习班，为研究生和博士后举办强化培训班。(3)此外，PI Baeumner将确保对本科生进行生物传感研究的培训，并继续与纽约州北部地区的印第安部落高中的高中生进行外联。