Biocal Technology, Inc.

Qiagen Sciences, LLC

Beckman Instruments, Inc.

Beckman Coulter, Inc.

Ming-Sun Liu

Varouj D Amirkhanian

Paul Mooney

Bob G Heitel

Andras Guttman

Fu-Tai A Chen

Ramon A Evangelista

Fu-Tai-A Chen

Sing-Tien Chiang

A cost-effective multi-channel capillary gel-electrophoresis system for highly efficient, nucleic acid-based analysis, in particular HLA SSP (Human Leukocyte Antigen Sequence Specific Primer) typing applications. Twelve DNA samples are automatically injected, electrophoretically separated, detected and analyzed simultaneously by using a multiple usage and disposable multi-capillary gel-cartridge. Using commercially available DNA size markers as indicators, the system provides high resolving power with 12-channel separations in less than 1.5 minutes. The system can hold a total of 96 samples in a PCR plate that can automatically be analyzed within 25 minutes for a full plate of the HLA SSP kits.

Method and apparatus for high speed genotyping

An integrated bio-analysis system incorporates built-in sample preparation capabilities. In one aspect of the present invention, a bio-analysis instrument is provided with a built-in sample preparation device based on PCR (or thermal cycling block/module). In one embodiment of the present invention, a peltier unit in the sample preparation device provides thermal cycling of samples supported in a multi-well tray. In another aspect of the present invention, a CE instrument is provided with a built-in sample preparation capability, which may comprise a sample preparation (bio-molecular reaction) device based on thermal cycler type. In another aspect of the present invention, a PCR device is provided with a built-in analysis device, such as a CE device, for verifying the results of the PCR (bio-molecular reaction) process.

Integrated bio-analysis and sample preparation system

A cost-effective multi-channel capillary gel-electrophoresis system for highly efficient, high speed, high throughput, carbohydrate analysis. In one aspect of the present invention, a high-performance capillary electrophoresis analyzer system has been optimized for carbohydrate analysis application. The system uses a multiplexed/time-staggered fluorescence type detection mechanism, with an integrated fiber optic array-based technology and a novel disposable gel-cartridge. Twelve carbohydrate samples are automatically injected, electrophoretically separated, detected and analyzed simultaneously by using a multiple usage and disposable multi-capillary gel-cartridge. Using commercially available labeling agent such as APTS (representing 8-Aminopyrene-1,3,6-trisulfonic acid, trisodium salt) as an indicator, the capillary electrophoresis-based carbohydrate system/analyzer provides high resolving power in 2-15 minutes of separations. The system can hold a total of 96 samples, which can be automatically analyzed within 30-60 minutes. This affordable fiber optic based fluorescence detection system can be used in laboratories for high speed carbohydrate sequencing & screening applications.

Method and apparatus for high speed carbohydrate analysis

A reusable bio-analysis cartridge that includes a built-in mechanism to track various parameters and usage data. The tracking mechanism includes a non-volatile rewritable memory 'smart-key' that is associated with the cartridge to provide automatic tracking dedicated to the cartridge. The key may be physically coupled to the cartridge with good high voltage insulating properties. When the cartridge is used in an instrument, the associated key is inserted into an I/O interface to communicate with the instrument. The instrument may be configured to 'authenticate' the cartridge and conduct an integrity check to determine if the particular cartridge has the correct properties (e.g., gel-chemistry, serial no., Patient I.D.) for the particular sample analysis to be conducted. Further, the instrument may communicate/record information concerning usage of the cartridge (e.g., usage history, test parameters, and perhaps test results). Such information provides an update to the stored information from the previous use of the cartridge.

Bio-analysis cartridge tracking and protection mechanism

The present invention provides for an interface mechanism in a bio-separation instrument that makes interface connections to a multi-channel cartridge. The interface mechanism precisely positions the cartridge in relation to the support elements in the instrument (e.g., high-voltage, gas pressure, incident radiation and detector), and makes automated, reliable and secured alignments and connections between various components in the cartridge and the support elements in the supporting instrument. The interface mechanism comprises pneumatically or electromechanically driven actuators for engaging support elements in the instrument to components on the cartridge. After the cartridge has been securely received by the interface mechanism, the connection sequence is initiated. The interface provides separate high voltage and optical connections for each separation channel in the cartridge, thus providing channel-to-channel isolation from cross talk both electrically and optically.

Multi-capillary electrophoresis cartridge interface mechanism

A bio-separation system using an efficient, compact, portable, interchangeable, reusable, recyclable, multi-channel cartridge, has integrated pre-aligned optics and an integrated reagent reservoir. The cartridge supports, for example, multiple capillaries for CE separation. An integrated reservoir containing a separation support medium (e.g., a gel buffer) is common to all capillaries. The chemistry of the medium and the characteristics of the capillaries (e.g., capillary size, coating and length) are defined for each cartridge. Different cartridges can be easily interchanged in the bio-separation system to suit the particular sample based separation. The reservoir is coupled to an air pressure pump that pressurizes the gel reservoir to purge and fill the capillaries with buffer as the separation support medium. In another aspect of the present invention, optics requiring fine alignment with respect to the detection zones (such as fiber optics for directing incident radiation and/or radiation emissions) are integrated into the cartridge.

Multi-channel bio-separation cartridge

A bio-separation instrument using a multi-segment cartridge. The cartridge includes a plurality of efficient, compact, portable, interchangeable, reusable, recyclable, modular, multi-channel segments. Each segment supports multiple capillaries for CE separation. Each segment has an integrated reservoir containing a buffer/gel common to all the capillaries. Air pressure can be supplied to the reservoir through the periphery of each segment. Each segment has an integrated LED array board directly coupled to capillaries for providing excitation light. Optical fibers of a fiber bundle are individually guided through the periphery of each segment to the capillaries for emission collection. The fiber bundle delivers emissions to a detector that can read the emissions in a time-staggered multiplexed scheme. Each segment includes electrodes electrically coupled to a voltage source through the periphery of the segment for effecting CE separation. Providing each segment with pneumatic and voltage connections on the periphery and direct coupling LED's allow the segments to easily stack together for integration into the instrument.

Multi-segment cartridge for bio-separation with multiplexed fluorescence detection

The invention provides novel electrophoretic methods for high-resolution separation and high-sensitivity detection of nucleic acids. The methods involve the use of a high-resolution buffer and a counter-migrating high-affinity intercalating dye in an electrophoresis system to achieve superior separation and detection of nucleic acids. A kit for separation and detection of nucleic acids in a sample by electrophoresis is also provided. The kit comprises a gel buffer containing a high-resolution intercalating dye with a resolution of at least 2.0 between the 271-bp and 282-bp &#x3c6;X 171 HAIII nucleic acid fragments nucleic acid fragments; and a high-affinity intercalating dye having a positive charge and DNA affinity constant of at least 10M.

Nucleic acid separation and detection by electrophoresis with a counter-migrating high-affinity intercalating dye

A multi-channel and multi-color bio-separation detection method and apparatus in which a single detector is coupled to a plurality of radiation sources, in a one detector/many radiation sources configuration. Each radiation source directs radiation at a detection zone of a single separation channel, and a single detector is applied to detect light emissions from the detection zones associated with several radiation sources. The radiation sources are activated to direct radiation at the detection zone in a predetermined sequence and further in a cyclic manner, with the detector output synchronized to the radiation sources by a controller. Bio-separation may be conducted simultaneously in all the channels in parallel, with detection time-staggered and/or time multiplexed with respect to the light sources. In one embodiment, low cost light emitting diodes may be used as radiation sources. In another aspect, the detection scheme is configured for radiation induced fluorescence detection in an capillary electrophoresis instrument.

Multi-color multiplexed analysis in a bio-separation system

A multi-channel bio-separation detection method and apparatus in which a single detector is coupled to a plurality of radiation sources, in a one detector/many radiation sources configuration. Each radiation source directs radiation at one detection zone of a single separation channel, and a single detector is applied to detect light emissions from the detection zones of several separation channels. The light sources direct radiation at the detection zones in a predetermined sequence and further in a cyclic manner, with the detector output synchronized to the light sources by a controller. Bio-separation may be conducted simultaneously in all the channels in parallel, with detection time-staggered and/or time multiplexed with respect to the light sources. In one embodiment, low cost light emitting diodes may be used as light sources. In another aspect, the detection scheme is configured for radiation induced fluorescence detection in a multi-channel capillary electrophoresis instrument.

Optical detection in a multi-channel bio-separation system

A process of collecting a sample of DNA from an individual for short-term or long term storage in the event that the DNA is required to identify the individual in the event of a cataclysmic accident. The collection of DNA sample from the individual begins with the completion of an application, for example for flight insurance, which is conveniently located at the airport, through a travel agent, etc. Upon completion of the application, a method of payment is determined and the DNA sample is collected. Preferably, the individual collects his or her DNA sample using a non-invasive approach such as swabbing the inside of his/her cheek and gum line. The DNA sample is then placed in a container and held in storage for a duration appropriate for the anticipated span of the activity or for a predetermined fixed term. At the conclusion of the activity, the DNA sample is either disposed of or returned to the individual for future use. If the individual is involved in an accident resulting in a cataclysmic death, the DNA is retrieved to aid in the identification of the deceased.

DNA sample collection for identification

In a bio-separation system, the detection zone for optical detection of sample analytes is located at a widened zone along the separation channel. In one embodiment of the present invention, the widened detection zone is a micro-bore collar having a micro-channel that coaxially surrounds the exit of a capillary column that defines a capillary channel. A separation support medium including a running buffer fills the capillary column and the collar. In another aspect of the present invention, incident radiation for detection of separated analytes is directed at the detection zone and/or emitted radiation is collected axially along the separation medium, instead of through the boundary walls of the detection zone. In a further aspect of the present invention, the optical detection configuration may be scaled up and implemented in a mult-channel CE system that comprises multiple capillary separation channels.

Optical detection in bio-separation device using a widened detection zone

Processes for the electrophoretic analysis of monosaccharides and/or polysaccharides are disclosed. The processes generally first include providing a composition having derivatized mono and/or polysaccharide components which are monosaccharide and/or polysaccharide labelled with a fluorescing charged compound. Then introducing the composition into an electrophoretic medium and applying an electric field across the electrophoretic medium causes the derivatized monosaccharide and/or polysaccharide components to differentially migrate within the electrophoretic medium. Then detecting the derivatized monosaccharide and/or polysaccharide components is accomplished by laser exciting the derivatized monosaccharide and/or polysaccharide components and monitoring fluorescent emission of the derivatized monosaccharide and/or polysaccharide components. The processes provide extremely low detection limits of the labelled components.

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Brea, CA, United States of America

Ming-Sun Liu