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12th International Symposium on Bioluminescence & Chemiluminescence |
Symposium abstracts:
Davies, Rupert H., Corry, Jacob W., Andrade, Joseph D.
Department of Bioengineering, University of Utah, Salt Lake City, Utah, USA
Email: rupert.davies@m.cc.utah.edu
A previously reported aqueous lactate assay, based on lactate dehydrogenase (LDH), flavin mononucleotide (FMN): reduced nicotinamide adenine dinucleotide (NADH) oxidoreductase, and bacterial luciferase, was enhanced with the additional of glutamic-alanine transaminase (GAT). The enhancement by GAT occurs via the consumption of pyruvate thereby increasing NADH production by LDH. This process was initially modeled and then assessed experimentally. This simple method of enhancement could be used in other bioluminescent-based assays. This aqueous assay was converted to a dry reagent assay with the addition of several excipients including sucrose, polyethylene glycol (PEG), dithiothreitol (DTT), bovine serum albumin (BSA), and ethylenediaminetetraacetic acid (EDTA). Formulations were tested before and after lyophilization as well as after days, weeks, and months of storage. Dry reagent assays are being miniaturized for the development of a multi-analyte biosensor.
This
is a preprint of an article accepted for publication in Luminescence: Copyright
2001 John
Wiley & Sons, Ltd (Wiley website)