Selective detection of thiamine using bacteria-derived proteins in complex biological matrices

Session: Thiamine Deficiency in the Great Lakes (1)

Katie Edwards, Binghamton University / Cornell , [email protected]
Eileen Randall, Cornell University, [email protected]
Esther Angert, Cornell University, [email protected]
Clifford Kraft, Cornell University, Dept. of Natural Resource, [email protected]

Abstract

Thiamine (vitamin B1) is essential to the health of nearly all living organisms, serving as an enzyme cofactor for numerous metabolic processes.  Deficiency in thiamine has had a marked impact on the health of salmonine fishes in the Great Lakes and other bodies of water, including the Finger Lakes and the Baltic Sea.  Early Mortality Syndrome, Cayuga Syndrome, and M74 syndrome all are related to thiamine deficiency and yield morbidity and mortality especially during developmental stages of fish. To improve the throughput and reduce costs associated with traditional HPLC-based thiamine analysis, we have developed a magnetic bead-based assay for thiamine that can be employed in a microtiter plate format. This assay relies on a protein sourced from  Escherichia coli  for thiamine recognition and has been demonstrated to have exceptional specificity towards thiamine versus its fragments or analogues. We will describe the development, performance relative to HPLC in salmonid eggs, liver, and muscle tissues, and benefits and caveats of this analytical approach.