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Stabilization of firefly luciferase activity with anti-oxidants: a preliminary study Rupert H. Davies*1; R Van Wagenen2; J Andrade1 (1) Bioengineering, Merrill Engineering Bld., 50 S. Central Campus Drive Rm. 2480, Salt Lake City, UT 84112, USA; (2) Protein Solutions, Inc.      *(rupert.davies@m.cc.utah.edu) Since biosensors are generally stored at room temperature for extended periods of time, the stability of firefly luciferase is essential for biosensor development. Lyophilization can increase firefly luciferase’s stability at room temperature; however, lyophilized firefly luciferase still loses activity with storage time. Some of this activity loss is likely due to oxidative stress. Oxidative stress caused by free radical species has been shown to significantly decrease the activity of firefly luciferase. Firefly luciferase activity depends on the presence of several sulfhydryl groups, which are especially susceptible to oxidation. Although DTT is generally used to protect the sulfhydryl groups, naturally occurring anti-oxidants such as vitamins C and E, beta-carotene, and glutathione have been shown to protect proteins against oxidative stress, individually and in concert. Therefore, the protection of firefly luciferase activity against oxidative stress by the naturally occurring anti-oxidants additives was studied and compared with DTT by using a typical ATP assay in aqueous, lyophilized, and immobilized forms. In addition, several anti-oxidants were studied in combination to mimic natural anti-oxidant systems. [Poster: davies.rupert.65742]

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