Enzymatic photochemical sensing using luciferase-immobilized polymer nanoparticles covered with artificial cell membrane.
We prepared phospholipid polymer nanoparticles immobilized with luciferase, and the nanoparticles were applied as photochemical sensing nanoparticles. An amphiphilic water-soluble polymer having a phosphorylcholine group was used as an emulsifier and a surface modifier to prepare the nanoparticles. The polymer was composed of three kinds of monomer units, that is, 2-methacryloyloxyethyl phosphorylcholine (MPC) as a hydrophilic and bioinert unit, n-butyl methacrylate as a hydrophobic unit and p-nitrophenyl ester having methacrylate as an enzyme-immobilizing unit. The p-nitrophenyl ester groups to immobilize the proteins were located on the surface of the nanoparticles. Luciferase was immobilized by the reaction between the p-nitrophenyl ester groups and the amino group. The enzymatic reaction on the nanoparticles was followed using a microdialysis system with an optical fiber having a 800 microm diameter in the probe. The nanoparticles conjugated with luciferase reacted with ATP, luciferin and oxygen. It is concluded that the nanoparticles are a promising tool for a photochemical sensing microdiagnostic system.
Konno
Tomohiro
T
Department of Materials Engineering, School of Engineering, The University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Ito
Tomomi
T
Takai
Madoka
M
Ishihara
Kazuhiko
K
D000255:Adenosine Triphosphate / Q000378:metabolism; D015374:Biosensing Techniques / Q000379:methods*; D004800:Enzymes, Immobilized; D049454:Firefly Luciferin / Q000378:metabolism; D008156:Luciferases / Q000378:metabolism*; D053758:Nanoparticles; D010100:Oxygen / Q000378:metabolism; D010777:Photochemistry; D011108:Polymers
D004800:Enzymes, Immobilized; D011108:Polymers; D049454:Firefly Luciferin; D000255:Adenosine Triphosphate; D008156:Luciferases; D010100:Oxygen