Use of restriction enzymes and other chemical methods to decrease non-specific binding in dual bead assays and related bio-discs, methods, and system apparatus for detecting medical targets

• Main Authors: YEUNG KAYUEN, LAM AMETHYST H, COOMBS JAMES H, VIRTANEN JORMA A, PHAN BRIGITTE C
• Format: Patent
• Language: English
• Published: 30.03.2006
• Subjects: ACTUATING-FLOATS; APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; BEER; BIOCHEMISTRY; BLASTING; CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE; CHEMISTRY; COCKS; COMPOSITIONS OR TEST PAPERS THEREFOR; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL ORENZYMOLOGICAL PROCESSES; DEVICES FOR VENTING OR AERATING; ENGINEERING ELEMENTS AND UNITS; ENZYMOLOGY; GENERAL MEASURES FOR PRODUCING AND MAINTAINING EFFECTIVEFUNCTIONING OF MACHINES OR INSTALLATIONS; HEATING; INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES; LIGHTING; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROM SOLIDMATERIALS OR FLUIDS; MAGNETIC OR ELECTROSTATIC SEPARATION OF SOLID MATERIALS FROMSOLID MATERIALS OR FLUIDS; MEASURING; MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEICACIDS OR MICROORGANISMS; MECHANICAL ENGINEERING; METALLURGY; MICROBIOLOGY; MUTATION OR GENETIC ENGINEERING; PERFORMING OPERATIONS; PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL; PHYSICS; PROCESSES OF PREPARING SUCH COMPOSITIONS; SEPARATION BY HIGH-VOLTAGE ELECTRIC FIELDS; SEPARATION OF SOLID MATERIALS USING LIQUIDS OR USING PNEUMATICTABLES OR JIGS; SPIRITS; TAPS; TESTING; THERMAL INSULATION IN GENERAL; TRANSPORTING; VALVES; VINEGAR; WEAPONS; WINE

Methods for decreasing non-specific bindings of beads in dual bead assays and related optical bio-discs and disc drive systems. Methods include identifying whether a target agent is present in a biological sample and mixing capture beads each having at least one transport probe affixed thereto, reporter beads each having at least one signal probe affixed thereto, and a biological sample. Mixing is performed under binding conditions to permit formation of a dual bead complex if the target agent is present in the sample. The reporter bead and capture bead each are bound to the target agent. Denaturing the target agent and keeping it in the denatured form by use of a specialized hybridization buffer is also provided. A denaturing agent is guanidine isothiocynate. Methods further include isolating the dual bead complex from the mixture to obtain an isolate, exposing the isolate to a capture field on a disc, and detecting the presence of the dual bead complex in the disc to indicate that the target agent is present in the sample. The methods may further include selectively breaking up non-specific binding between capture beads and reporter beads employing a digestion agent. Also employed is a method for selectively breaking up non-specific binding between capture beads and reporter beads using a wash buffer containing a chemical agent. The methods are applied to detecting medical targets.