An enzyme-immobilization method for integration of biofunctions on a microchip using a water-soluble amphiphilic phospholipid polymer having a reacting group.
A water-soluble phospholipid polymer having an active ester group in the side chain, poly[2-methacryloyloxyethyl phosphorylcholine (MPC)-co-n-butyl methacrylate (BMA)-co-p-nitrophenyloxycarbonyl polyethyleneglycol methacrylate (MEONP)(PMBN), was used for the immobilization of an enzyme on a plastic microchip. The MPC polymers with BMA units were adsorbed onto the poly(methyl methacrylate)(PMMA) microchip, and the active ester group in the MEONP unit reacted with the amino groups of the proteolytic enzyme, trypsin. Trypsin was immobilized on the sample reservoir, and catalyzed the hydrolysis of the fluorescently labeled ArgOEt to Arg. The consequent separation of product from the substrate, and their detection, were integrated on the microchip and this meant that all procedures from the enzymatic activity to product detection were completed in less than three minutes.
Sakai-Kato
Kumiko
K
Department of Analytical Chemistry, School of Pharmaceutical Sciences, University of Shizuoka, Shizuoka, Japan. daikato@u-shizuoka-ken.ac.jp
Kato
Masaru
M
Ishihara
Kazuhiko
K
Toyo'oka
Toshimasa
T
D000327:Adsorption; D001120:Arginine / Q000031:analogs & derivatives / Q000737:chemistry; D004800:Enzymes, Immobilized / Q000737:chemistry*; D005456:Fluorescent Dyes; D006868:Hydrolysis; D015394:Molecular Structure; D010743:Phospholipids / Q000737:chemistry*; D010767:Phosphorylcholine; D019904:Polymethyl Methacrylate / Q000737:chemistry*; D013997:Time Factors; D014357:Trypsin / Q000737:chemistry; D014867:Water / Q000737:chemistry
D004800:Enzymes, Immobilized; D005456:Fluorescent Dyes; D010743:Phospholipids; D014867:Water; D010767:Phosphorylcholine; D019904:Polymethyl Methacrylate; D001120:Arginine; C489258:arginine ethyl ester; D014357:Trypsin