Luciferase firefly, biotinylated

Biotinylated antibodies can via streptavidin be linked to biotinylated enzymes, e.g. firefly luciferase or an ATP producing enzyme like acetate kinase. A genetically fused biotinylated thermostable recombinant luciferase can be detected below attomol levels and is used in a kit for the detection of 10 cfu/mL of Staphylococcus aureus. In another kit coliforms are detected by their P-galactosidase activity using D-luciferin-O-P-galactopyranoside and the firefly reaction. ... 

Fukoda S, Tatsumi H, Maeda M. Bioluminescent enzyme immunoassay with biotinylated firefly luciferase. J Clin Ligand Assay 1998 21 358-362. 

Tatsumi H, Fukuda S, Kikuchi M, Koyama Y. Construction of biotinylated firefly luciferase using biotin acceptor peptides. Anal Biochem 1996 243 176-80. 

Reagents. Recombinant aequorin (Aq) with a free cys residue was from Chisso Corporation (Yokohama, Japan). Thermostable biotinylated firefly luciferase (b-Luc) was obtained from Kikkoman Corporation (Chiba, Japan). Aq labeled anti-Dig Fab fragment and b-Luc/streptavidin complex were produced by previously reported.1 HRP-labeled anti-DNP was purchased from LSL Co. (Tokyo). 

Simultaneous luminescent measurement of aequorin, biotinylated firefly luciferase and HRP. Aq, b-Luc and HRP solutions (5 pL of each) diluted with 0.05 mol/L HEPES-KOH buffer (pH 7.0) containing 5 mmol/L EGTA and 10% Block Ace (Dainippon SumitomoPharma Co., Ltd., Osaka, Japan) were introduced... 

Ito K, Nishimura W, Maeda M, et al. Highly sensitive and rapid tandem bioluminescent immunoassay using aequorin labeled Fab fragment and biotinylated firefly luciferase. Anal Chim Acta 2007 588 245-51. 

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