Apparatus for active biological sample preparation

• US 6,824,740 B1
• Filed: 04/28/2000
• Issued: 11/30/2004
• Est. Priority Date: 09/06/1996
• Status: Expired due to Fees

• Current Assignee: Gamida for Life B.V.
• Original Assignee: Nanogen, Inc.
• Inventors: Jackson, Thomas R., Scott, Bradley S., Sheldon, Edward L. III, Heller, Michael J., Swanson, Paul D.
• Primary Examiner(s): Nguyen, Nam
• Assistant Examiner(s): STARSIAK, JOHN S
• Application Number:
  US09/561,318
• Time in Patent Office: 1,677 Days
• Field of Search: 204/518, 204/542, 204/543, 204/544, 204/627, 204/641, 204/604, 204/607, 204/608, 435/287.2, 435/288.2, 435/287.3, 435/288.3, 435/288.4, 435/288.5, 435/288.6, 435/288.7, 422/99, 422/100
• US Class Current: 422/527
• CPC Class Codes: B01D 57/02 by electrophoresis treatment of water, waste water, sewage or sludge by electrophoresis C02F1/469; electrophoretic production of compounds or non-metals C25B7/00; investigating or analysing materials by using electrophoresis G01N27/26
• CPC Class Codes: B01J 19/0093 Microreactors, e.g. miniaturised or microfabricated reactors laboratory containers with capillary fluid transport in microfabricated channels or chambers B01L3/5027
• CPC Class Codes: B01L 2200/0631 Purification arrangements, e.g. solid phase extraction [SPE]
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2300/0636 Integrated biosensor, microarrays
• CPC Class Codes: B01L 2300/0645 Electrodes
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 2400/0421 electrophoretic flow
• CPC Class Codes: B01L 2400/0424 Dielectrophoretic forces
• CPC Class Codes: B01L 2400/0644 rotary valves
• CPC Class Codes: B01L 3/502753 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• CPC Class Codes: B03C 5/026 using open-gradient differential dielectric separation, i.e. using electrodes of special shapes for non-uniform field creation, e.g. Fluid Integrated Circuit [FIC]
• CPC Class Codes: B03C 5/028 using travelling electric fields, i.e. travelling wave dielectrophoresis [TWD]
• CPC Class Codes: C12N 13/00 Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
• CPC Class Codes: G01N 27/44743 Introducing samples
• CPC Class Codes: H01L 29/6656 using multiple spacer layers, e.g. multiple sidewall spacers

Abstract

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.

Claims

1. A system for the active biological sample preparation of materials comprising:

a sample addition port,

a purification chamber, the purification chamber being positioned downstream of the sample addition port, and including at least a first electrophoretic solution, and first and second electrophoresis electrodes and a membrane at one end of the purification chamber,

a protein trap within the first electrophoretic solution,

a tap connected to the purification chamber,

a denaturation chamber positioned downstream of the tap,

a complexity reduction chamber positioned downstream of the denaturation chamber, and

a diagnostic chamber positioned downstream of the complexity reduction chamber.

2. The system of claim 1 further including a cell lysing device disposed between the sample addition port and the purification chamber.

3. The apparatus of claim 1 further including a third electrode, the third electrode located adjacent the tap and adapted to move materials from the purification chamber to the tap.

4. The apparatus of claim 3 wherein the electrode is a C shaped electrode.

5. The apparatus of claim 3 wherein the third electrode includes:

first and second portions generally disposed in a direction which is oblique to the axis defined by the first and second electrodes, and a third portion positioned for driving material towards the tap.

6. The system of claim 1 further comprising a reagent causing cell membrane disruption disposed between the sample addition port and the purification chamber.