Nanogen, Inc.

Affymetrix, Inc.

Molecular Devices Corporation

Affymax Technologies, N.v.

Gamida for Life B.v.

Edward L Sheldon

Mark S Chee

Earl A Hubbell

Robert J Lipshutz

Maureen T Cronin

Peter E Lobban

Donald E Ackley

Charles Garrett Miyada

Xiaohua C Huang

Stephen P A Fodor

MacDonald S Morris

Jing Cheng

Thomas R Jackson

Lei Wu

Viola T Kung

James P O'Connell

Michael K Krihak

Michael James Heller

Stephen P Fodor

Paul David Swanson

Robert F Zuk

John D Olson

Macdonald S Morris

Bradley S Scott

Peter A Nagainis

Richard D Armenta

Charles R Burke

Glenn H McGall

Thomas R Gingeras

Jonathan M Diver

Dan Smolko

Shila Jalali

David Willoughby

Xiaohua X Huang

Nila Shah

We have performed separation of bacterial and cancer cells from peripheral human blood in microfabricated electronic chips by dielectrophoresis. The isolated cells were examined by staining the nuclei with fluorescent dye followed by laser induced fluorescence imaging. We have also released DNA and RNA from the isolated cells electronically and detected specific marker sequences by DNA amplification followed by electronic hybridization to immobilized capture probes. Efforts towards the construction of a 'laboratory-on-a-chip' system are presented which involves the selection of DNA probes, dyes, reagents and prototyping of the fully integrated portable instrument.

Integrated portable biological detection system

The invention provides arrays of immobilized probes, and methods employing the arrays, for detecting mutations in the biotransformation genes, such as cytochromes P450. For example, one such array comprises four probe sets. A first probe set comprises a plurality of probes, each probe comprising a segment of at least three nucleotides exactly complementary to a subsequence of a reference sequence from a biotransformation gene, the segment including at least one interrogation position complementary to a corresponding nucleotide in the reference sequence. Second, third and fourth probe sets each comprise a corresponding probe for each probe in the first probe set. The probes in the second, third and fourth probe sets are identical to a sequence comprising the corresponding probe from the first probe set or a subsequence of at least three nucleotides thereof that includes the at least one interrogation position, except that the at least one interrogation position is occupied by a different nucleotide in each of the four corresponding probes from the four probe sets.

Arrays of nucleic acid probes for analyzing biotransformation genes

The invention provides methods of comparing a target nucleic acid with a reference nucleic acid using nucleic acid arrays.

Method of comparing a target nucleic acid and a reference nucleic acid

The invention provides chips of immobilized probes, and methods employing the chips, for comparing a reference polynucleotide sequence of known sequence with a target sequence showing substantial similarity with the reference sequence, but differing in the presence of e.g., mutations.

Arrays of nucleic acid probes on biological chips

The present invention comprises devices and methods for performing channel-less separation of cell particles by dielectrophoresis, DC high voltage-pulsed electronic lysis of separated cells, separation of desired components from crude mixtures such as cell lysates, and/or enzymatic reaction of such lysates, all of which can be conducted on a single bioelectronic chip. A preferred embodiment of the present invention comprises a cartridge () including a microfabricated silicon chip () on a printed circuit board () and a flow cell () mounted to the chip () to form a flow chamber. The cartridge () also includes output pins () for electronically connecting the cartridge () to an electronic controller. The chip () % includes a plurality of circular microelectrodes () which are preferably coated with a protective permeation layer. Specific cells from various cell mixtures were separated, lysed, and enzymatically digested on the chip.

Channel-less separation of bioparticles on a bioelectronic chip by dielectrophoresis

Systems and methods for the electronic sample preparation of biological materials utilize the differential charge-to-mass ratio and/or the differential affinity of sample constituents to separation materials for sample preparation. An integrated system is provided for performing some or all of the processes of: receipt of biological materials, cell selection, sample purification, sample concentration, buffer exchange, complexity reduction and/or diagnosis and analysis. In one embodiment, one or more sample chambers adapted to receive a buffer solution are formed adjacent to a spacer region which may include a trap or other affinity material, electrophoretic motion of the materials to be prepared being effected through operation of electrodes. In another aspect of this invention, a transporter or dipstick serves to collect and permit transport of materials, such as nucleic acids, most preferably DNA and/or RNA. In one embodiment, a membrane or trap is held in a frame which is adapted to mate with a channel formed in the spacer region. In another aspect of this invention, an electrophoretic system for biological sample preparation is operated in a manner so as to utilize the differential charge-to-mass ratio so as to control the migration of materials within the solution. In one aspect, bunching of selected materials is achieved by operation of two electrodes in a manner so as to reduce the spatial dispersion of those materials. In another aspect of this invention, a vertically disposed sample preparation unit includes an upper reservoir including and a collection chamber. A sample is preferably pre-prepared and densified, applied to the conductive polymer, electrophoresed so as to move nucleic acids into the conductive polymer and move undesired material away from the conductive polymer. Integrated systems are described in which cell separation, purification, complexity reduction and diagnosis may be performed together. In the preferred embodiment, cell separation and sample purification are performed in a first region, the steps of denaturation, complexity reduction and diagnosis being performed in a second region.

Apparatus for active biological sample preparation

Methods and apparatus for use of a stacked, reconfigurable system is provided. The stacked, reconfigurable system includes an inlet for receipt of a sample, a first chamber defined by a bottom support, an intermediate member, and a first spacer, the first chamber being coupled to the inlet through the bottom support, the first chamber including an analysis system having electrodes for electrophoretic transport of material. A second chamber is stacked on top of the first chamber, the second chamber being defined by the intermediate member, a top member, and a second spacer, the second chamber being in fluid communication with the first chamber through at least one via formed in the intermediate member, the second chamber being electrically reconfigurable to permit action such as, for example, synthesis of a compound or specialized analysis. The apparatus can be used to perform analysis on charged biological materials.

Reconfigurable detection and analysis apparatus and method

The invention provides methods for detecting variations in polymorphic sites and/or variations in gene copy number. The methods are particularly useful for analysis of biotransformation genes, such as cytochromes P450.

Analysis of genetic polymorphisms and gene copy number

A multilayer, laminated device for performing fluidic operations includes a first exterior support layer, an adjacent layer disposed in laminated relationship with a portion of the exterior support layer, a second exterior support layer disposed in laminated relationship with a portion of the adjacent layer, and a fluidic pump that is disposed in the adjacent layer. The fluidic pump includes a first gear rotationally mounted to the adjacent layer, the first gear having a magnet contained therein. A second gear is rotationally mounted to the adjacent layer, the second gear having a magnet contained therein, the second gear being engaged with the first gear in a meshed relationship.

Laminated assembly for active bioelectronic devices

Methods, apparatus, and applications for use of a stacked, reconfigurable system for electrophoretic transport are provided. In one embodiment, a system having a first chamber including at least a bottom support and an intermediate support, and a second chamber, said second chamber including a bottom support and a top member, the first and second chambers being coupled through a via. Electrophoretic, and optional electro-osmotic and thermal, transport is effected. In another aspect of this invention, three or more chambers are coupled by an electrophoretic buss. The electrophoretic buss includes driving electrodes and is adapted to receive fluid containing materials for transport. The chambers are coupled to the electrophoretic buss and serve as a tap from the buss for delivery of charged materials. In one embodiment, certain functions are performed in different chambers. For example, the first chamber may receive the sample and perform sample processing functions, the second chamber may perform amplification procedures, yet a third chamber may perform hybridization or other assays, and yet another chamber may perform immunoassays. By separating various functions to different chambers, speed and sensitivity may be improved. In yet another aspect of this invention, analysis from a earlier stage may be utilized in a subsequent stage to reconfigure the system for optimum use. In one application, analysis at a first level is utilized to determine an action at a second level, such as the synthesis of a compound. The synthesized compound in response to a biohazard may comprise vaccine or antidote.

Electrophoretic buss for transport of charged materials in a multi-chamber system

System including functionally separated regions in electrophoretic system

The invention provides arrays of immobilized probes, and methods employing the arrays, for detecting mutations in the biotransformation genes, such as cytochromes P450. For example, one such array comprises four probe sets. A first probe set comprises a plurality of probes, each probe comprising a segment of at least three nucleotides exactly complementary to a subsequence of a reference sequence from a biotransformation gene, the segment including at least one interrogation position complementary to a corresponding nucleotide in the reference sequence. Second, third and fourth probe sets each comprise a corresponding probe for each probe in the first probe set. The probes in the second, third and fourth probe sets are identical to a sequence comprising the corresponding probe from the first probe set or a subsequece of at least three nucleotides thereof that includes the at least one interrogation position, except that the at least one interrogation position is occupied by a different nucleotide in each of the four corresponding probes from the four probe sets.

Arrays of nucleic acid probes for analyzing biotransformation genes and methods of using the same

Stacked, reconfigurable system for electrophoretic transport of charged materials

The present invention encompasses a capture membrane comprising a porous filter membrane having a hapten bound directly or indirectly to the membrane wherein complexes formed by specific binding having an anti-hapten bound to a binding member of the specifically binding complex are removed from a solution by the hapten as the solution passes through the membrane. In the preferred embodiment biotin is the hapten and avidin or streptavidin is the anti-hapten.

Hapten derivatized capture membrane and diagnostic assays using such membrane

Methods of manufacture and devices for performing active biological operations utilize laminated structures. In the preferred embodiment, a first planar sample support includes at least one sample through hole, a planar electrode is disposed adjacent the first planar sample support, and includes an electrode through region, a second planar support includes a vent through hole, the planar electrode being in a laminated relationship between the first planar sample support and the second planar support, further characterized in that the sample through hole, electrode through hole and vent through hole are in overlapping arrangement. Preferably, some or all of the through holes, through regions and vent through holes are aligned. In one embodiment, the lateral dimension of the vent through hole is larger than the lateral dimension of the electrode through hole. In an alternative embodiment, the lateral dimension of the sample through hole is larger than the lateral dimension of the vent through hole.

Growing community of inventors

San Diego, CA, United States of America

Edward L Sheldon