BIOMONAR: Biosensor nanoarrays for environmental monitoring

1064267 Baeumner环境样品是复杂的，为了检测环境分析物，以“一刀切”的生物传感器为目标是不现实的。与此同时，非系统的生物传感器设计方法导致了一套非常多样化的传感器，几乎没有共同特征，阻碍了对环境过程和新目标传感器设计的必要的概念性和基础性的理解。因此，一组欧洲研究人员和Pi Baeumner在2010年得到欧盟委员会的资助(为期4年)，用于开发用于测量一系列化学和生物参数的纳米阵列生物传感器电池。新型生物识别元件的开发和基础研究，对目标分析物的反应性和通量的研究，为设计复杂环境样品中新分析物的传感器提供了基本的理解。具体地说，重点是开发三个互补的传感平台(光学、脂质体、全细胞)，将单个传感蛋白家族整合为生物识别元件，能够传感无限数量的污染物。细菌周质结合蛋白(PBPs)是一种基因工程技术，可与多种与环境分析有关的分析物(包括重金属、农药和病原体)特异和灵敏地结合。为了在通过欧盟项目应用基于脂质体的PBP传感平台之前对其有基本的了解：(1)利用表面等离子共振(SPR)对PBPs作为生物识别元件有基本的了解，包括亲和力和动力学结合研究；(2)发展基于脂质体的PBP微孔板荧光分析方法，通过对蛋白质的量化表征，获得关于PBP作为环境基质中的生物识别元件的信息，(3)开发基于电化学发光的微流控脂质体生物传感器，用于环境分析，避免与基质相关的非特异性信号。这是与欧盟合作伙伴开展的研究的补充，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和PBP分析物结合强度的测定，(5)确定脂质体增强Mach-Zehnder干涉仪灵敏度的能力的研究，以及(6)将脂质体作为水凝胶中的细胞模拟物的研究，以帮助研究无机和有机化合物的生物利用度。此外，还将通过基于脂质体的电化学发光(ECL)微流控策略开发高灵敏度和高分辨率的环境生物传感器。此外，通过与欧盟伙伴的密集互动，脂质体将被作为多功能颗粒进行研究。欧盟伙伴和Pi Baeumner的整体研究的科学价值基于多学科方法，该方法开发了一个通用动态框架，用于定量解释确定污染物生物影响的“暴露于效应”的过程链。更广泛的影响包括(1)欧盟团队内商业环境监测机构合作伙伴建造和测试用于环境监测的新一代生物传感器。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，可能有助于制定环境政策。(2)为最终用户组织讲习班，并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方面的培训，并继续与纽约州北部地区的一所印第安人部落高中的高中生进行接触。

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.