Ultrasensitive ATP detection using firefly luciferase entrapped in sugar-modified sol-gel-derived silica.
Firefly luciferase (FL) was entrapped in sol-gel-derived silica containing precursors based on covalent linkage of d-gluconolactone or d-maltonolactone to (aminopropyl)triethoxysilane to form N-(3-triethoxysilylpropyl)gluconamide or N-(3-triethoxysilylpropyl)maltonamide. The enzyme was active and stable in this material and showed catalytic constants close to those in solution. As little as 20 amol ATP could be detected with the entrapped FL, and the entrapped enzyme could be used over several cycles.
Cruz-Aguado
Jorge A
JA
Department of Chemistry, McMaster University, 1280 Main Street West, Hamilton ON, Canada L8S 4M1.
Chen
Yang
Y
Zhang
Zheng
Z
Elowe
Nadine H
NH
Brook
Michael A
MA
Brennan
John D
JD
D000255:Adenosine Triphosphate / Q000032:analysis*; D000818:Animals; D002241:Carbohydrates / Q000737:chemistry*; D002384:Catalysis; D001517:Coleoptera / Q000201:enzymology*; D005782:Gels / Q000737:chemistry*; D008156:Luciferases / Q000378:metabolism*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012822:Silicon Dioxide / Q000737:chemistry*; D013050:Spectrometry, Fluorescence
D002241:Carbohydrates; D005782:Gels; D012822:Silicon Dioxide; D000255:Adenosine Triphosphate; D008156:Luciferases