title: Immobilization of aptamer-based molecular beacons onto optically-encoded micro-sized beads.
abstract: This paper presents a method for the novel immobilization of aptamer-based molecular beacons (apta-beacons) onto optically-encoded micro-sized beads (apta-beacon beads). To immobilize apta-beacons onto flourescently-encoded micro-sized beads, core-shell type beads containing a fluorescent dye-encoded core and apta beacon-coupled shell were prepared. The fluorescent dye-encoded beads were prepared from TentaGel resins by coupling RITC to the amino groups of the core region, after partial protection of amino groups with Fmoc-OSu in a diffusion-controlled manner. After deprotection of the Fmoc-amino groups, FITC-coupled molecular beacons (MBs) were immobilized to the beads together with a quencher by covelent bonding. Briefly, aspartic acid (Asp) was coupled to the shell part of the beads. Then, the quencher was coupled to the N-terminal amino group of Asp and the MBs were coupled to the side chain carboxyl group. In a model study, thrombin was directly detected using this apta-beacon bead method. The thrombin-bound apta-beacon beads were easily recognized by the appearance of fluorescence without any further labeling step.
authors: School of Chemical and biological Engineering, Seoul National University, Seoul 151-747, Korea.
authors: Bong-Hyun
authors: BH
authors: Jun
authors: Ji-Eun
authors: JE
authors: Kim
authors: Chul
authors: C
authors: Rho
authors: Jang-Woong
authors: JW
authors: Byun
authors: Yo Han
authors: YH
authors: Kim
authors: Homan
authors: H
authors: Kang
authors: Jong-Ho
authors: JH
authors: Kim
authors: Taegyu
authors: T
authors: Kang
authors: Myung-Haing
authors: MH
authors: Cho
authors: Yoon-Sik
authors: YS
authors: Lee
mesh_terms: D052158:Aptamers, Peptide / Q000737:chemistry*; D001224:Aspartic Acid / Q000737:chemistry; D015374:Biosensing Techniques / Q000295:instrumentation*; D005449:Fluorenes / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D057166:High-Throughput Screening Assays; D055767:Immobilized Proteins / Q000737:chemistry*; D008863:Microspheres; D008967:Molecular Biology / Q000379:methods*; D036103:Nanotechnology / Q000295:instrumentation*; D012235:Rhodamines / Q000737:chemistry; D013917:Thrombin / Q000737:chemistry
chemical_list: D052158:Aptamers, Peptide; D005449:Fluorenes; D005456:Fluorescent Dyes; D055767:Immobilized Proteins; D012235:Rhodamines; D001224:Aspartic Acid; D013917:Thrombin