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

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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.