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

PI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。一种基于免疫亲和固相萃取的流动注射免疫传感器可以通过远程控制技术或在船上对海水中的超痕量B12进行快速原位测量。一种基于上覆水方法优化的荧光共振能量转移（FRET）平面免疫传感器将用于底部沉积物中维生素B12的二维分布和运输模式的原位测绘。维生素B12是水生系统中浮游植物初级生产所特别需要的微生物产生的化合物。它是生物生长和功能所必需的多种微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才刚刚开始被认识，由于缺乏合适的分析工具，海洋沉积物中B12和其他生物源有机金属的分布、运输和控制生产模式的因素基本上没有记载。对维生素B12进行快速、准确的原位定量或定性侦察的能力将大大提高对维生素及其相关化合物的生物地球化学循环的认识，进一步揭示维生素B12对初级生产力和浮游植物生长的影响，并为沿海环境中营养循环的概念模型和实际管理决策提供增强的基础。π吗?i’申请资助，开发利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维生素B12的ELISA方法。一种基于免疫亲和固相萃取的流动注射免疫传感器可以通过远程控制技术或在船上对海水中的超痕量B12进行快速原位测量。一种基于上覆水方法优化的荧光共振能量转移（FRET）平面免疫传感器将用于底部沉积物中维生素B12的二维分布和运输模式的原位测绘。更广泛的影响：这项提议的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有重要的应用，比如化学生态学。该项目有潜力开发一种新型传感器，如果成功，将大大有助于我们了解B12对海洋系统的影响。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课程，以及当地高中的推广项目，以鼓励积极进取的学生。

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.