Authors: W.C. Maki, N.N. Mishra, B. Filanoski, S.K. Rastogi, E. Cameron, G. Maki
Affilation: University of Idaho, United States
Pages: 248 - 251
Keywords: silica nano-particle, CMOS chip, Staphylocuccus aureus, luciferase
Mesoporous silica nano-particles were immobilized with luciferase and antibodies for immune-bioluminescence detection of Staphylocuccus aureus enterotoxin B (SEB) and C (SEC). Mesoporous silica nano-particles were used as matrix to immobilize antibodies specific for enterotoxins and luciferases from the Jamaican click beetles which were used as reporters. Fluorescent labeled mouse IgG and green fluorescent protein (GFP) were used to test and confirm the protein immobilization on the nano-particles by fluorescent microscopic method. For the application of functionalized meso-porous bio-luminescence nanoparticle in bio-molecular detection, two sets of nano-particles were prepared: anti-SEB antibody with luciferase-100 (red emission) and anti-SEC antibody with luciferase-99 (green emission). Antibody-antigen recognition was performed in 96 assay plates and on nitrocellulose membrane. In the presence of the target molecules, the enzymatic reaction on the nanoparticles was detected by using luminescence detector and CCD camera. We were able to preserve the enzyme activity and antibody biological function on these nanoparticles for longer period of time. We also demonstrated the detection of this bioluminescence reaction on a simple CMOS imaging chip based device.