Immunosensors for water column and 2-D sediment distributions of vitamin B-12 and target organic solutes

PI请求资助开发ELISA方法，使用稳定和时间分辨荧光技术测定沿海海水和沉积物孔隙水中的维生素B12。基于免疫亲和固相萃取的流动注射免疫传感器将允许通过远程控制技术或在船上快速原位测量海水中的超痕量B12。一种新的荧光共振能量转移（FRET）平面免疫传感器，优化的上覆水的方法的基础上，将被用于原位映射的二维维生素B12的分布和运输模式在底部沉积物。 维生素B12是一种微生物产生的化合物，特别是浮游植物在水生系统中初级生产所需的化合物。它是生物生长和功能所必需的多种痕量有机金属化合物的代表，主要来源于沉积物中的异养微生物群落。它在浮游植物生态学中的重要性刚刚开始被理解，由于缺乏合适的分析工具，B12和其他生物有机金属在海洋沉积物中的分布，运输和控制生产模式的因素基本上没有记录。对维生素B12进行快速、准确的定量或定性调查的能力将大大增进对维生素和相关化合物的生物地球化学循环的了解，进一步揭示B12对初级生产力和浮游植物生长的影响，并为沿海环境中与营养物循环有关的概念模型和实际管理决策提供更好的基础。 私家侦探？2009年，美国申请资助开发ELISA方法，使用稳定和时间分辨荧光技术测定沿海海水和沉积物孔隙水中的维生素B12。基于免疫亲和固相萃取的流动注射免疫传感器将允许通过远程控制技术或在船上快速原位测量海水中的超痕量B12。一种新的荧光共振能量转移（FRET）平面免疫传感器，优化的上覆水的方法的基础上，将被用于原位映射的二维维生素B12的分布和运输模式在底部沉积物。 更广泛的影响： 这项工作可能会彻底改变我们分析小生物分子的方式，并可能在化学海洋学以外的领域（如化学生态学）中有重要的应用。 该项目有可能开发一种新型传感器，如果成功，将大大有助于我们了解B12在海洋系统中的影响。这些新技术（流动免疫传感器，平面荧光）将被纳入研究生和本科生课程，以及当地高中推广计划，以高度积极的学生。

The PI's request funding to develop ELISA methods for determination of vitamin B12 in coastal seawater and sediment pore water using both steady and time-resolved fluorescence technologies. A flow injection immunosensor based on immunoaffinity solid-phase extraction will allow rapid in-situ measurement of ultra trace B12 in seawater either by remote control technology, or on board ship. A new fluorescence resonance energy transfer (FRET) planar immunosensor, optimized based on the overlying water methodology, will be utilized for in-situ mapping of 2-dimensional vitamin B12 distribution and transport patterns in bottom deposits. Vitamin B12 is a microbially produced compound specifically required by phytoplankton for primary production in aquatic systems. It is representative of multiple trace organometallic compounds essential for biological growth and functioning, and has major sources from heterotrophic microbial communities in sediments. Its importance in phytoplankton ecology is just beginning to be understood, and the distribution, transport and factors controlling production patterns of B12 and other biogenic organometallics in marine sediments are essentially undocumented due to lack of suitable analytical tools. The capability for rapid and accurate quantitative or qualitative reconnaissance of vitamin B12 in situ would significantly advance knowledge of biogeochemical cycling of vitamins and related compounds, further reveal the influence of B12 on primary productivity and phytoplankton growth, and provide an enhanced basis for conceptual models and practical management decisions related to nutrient cycling in coastal environments. The PI?s request funding to develop ELISA methods for determination of vitamin B12 in coastal seawater and sediment pore water using both steady and time-resolved fluorescence technologies. A flow injection immunosensor based on immunoaffinity solid-phase extraction will allow rapid in-situ measurement of ultra trace B12 in seawater either by remote control technology, or on board ship. A new fluorescence resonance energy transfer (FRET) planar immunosensor, optimized based on the overlying water methodology, will be utilized for in-situ mapping of 2-dimensional vitamin B12 distribution and transport patterns in bottom deposits. Broader Impacts: The proposed work could revolutionize the way we analyze small biomolecules, and may have important applications in fields outside of chemical oceanography, like chemical ecology. This project has the potential develop a novel sensor that if successful would contribute significantly to our understanding of the influence of B12 in marine systems. These novel technologies (flow immunosensor, planar fluorescence) would be incorporated into graduate and undergraduate curricula, as well to local high school outreach programs to highly motivated students.