Methods and apparatus for processing a sample of biomolecular analyte using a microfabricated device

• US 6,207,031 B1
• Filed: 09/14/1998
• Issued: 03/27/2001
• Est. Priority Date: 09/15/1997
• Status: Expired due to Term

• Current Assignee: Whitehead Institute for Biomedical Research
• Original Assignee: Whitehead Institute for Biomedical Research
• Inventors: Adourian, Aram S., Schmalzing, Dieter R., Matsudaira, Paul T., Ehrlich, Daniel J., Koutny, Lance B.
• Primary Examiner(s): Soderquist, Arlen
• Assistant Examiner(s): STARSIAK, JOHN S
• Application Number:
  US09/153,215
• Time in Patent Office: 925 Days
• Field of Search: 204/600, 204/601, 204/602, 204/603, 204/604, 204/605, 204/606, 204/615, 204/616, 204/617, 204/618, 204/619, 204/620, 204/450, 204/451, 204/452, 204/453, 204/455, 204/456, 204/461, 204/466, 204/467, 422/100, 422/63, 422/67
• US Class Current: 204/451
• CPC Class Codes: B01L 2200/027 for microfluidic devices
• CPC Class Codes: B01L 2200/0605 Metering of fluids
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/0657 Pipetting powder
• CPC Class Codes: B01L 2300/0803 Disc shape
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/0421 electrophoretic flow
• CPC Class Codes: B01L 2400/043 magnetic forces
• CPC Class Codes: B01L 2400/0442 thermal energy, e.g. vaporisation, bubble jet
• CPC Class Codes: B01L 3/0275 Interchangeable or disposable dispensing tips
• CPC Class Codes: B01L 3/0289 Apparatus for withdrawing or distributing predetermined quantities of fluid B01L3/02 takes precedence; sample taking G01N1/00; sample taking within automatic analysers G01N35/00; volume measuring in general G01F
• CPC Class Codes: B01L 3/5025 for parallel transport of multiple samples
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/502715 characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
• 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: G01N 27/44743 Introducing samples
• CPC Class Codes: G01N 27/44782 of a plurality of samples
• CPC Class Codes: G01N 35/10 Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices G01N35/0099 takes precedence
• CPC Class Codes: Y10T 436/2575 Volumetric liquid transfer

Abstract

A technique processes a sample of biomolecular analyte. The technique uses an apparatus having a support assembly that receives and supports a test module, a load assembly that loads the sample of biomolecular analyte onto the test module, an electrophoresis assembly that applies a current to the test module such that components within the sample separate by electrophoresis, and a controller that controls operations of the load assembly and the electrophoresis assembly. The load assembly and the electrophoresis assembly are coupled to the support assembly. The controller controls the operation of the load assembly in an automated manner. Preferably, the test module includes a dielectric plate member having an upper planar surface and a lower planar surface that is spaced apart from and coplanar with the upper planar surface. The dielectric plate member has at least one set of channels that includes an injection channel and a separation channel. The injection channel extends from the upper planar surface to the lower planar surface. The separation channel extends within the dielectric plate member in a plane parallel with the upper and lower planar surfaces and intersects the injection channel.

Claims

1. An apparatus for processing a sample of biomolecular analyte, comprising:

a support assembly that receives and supports at least one test module having micromachined separation channels optimized for high throughput processing;

a load assembly, coupled to the support assembly, that loads the sample of biomolecular analyte onto the test module;

an electrophoresis power assembly, coupled to the support assembly, that applies a current to the test module such that components within the sample separate by electrophoresis in the micromachined separation channels, the micromachined separation channels defining micron amounts of the sample such that electrophoresis at a high throughput rate is enabled; and

a controller, adjacent the support assembly, that controls operations of the load assembly and the electrophoresis power assembly, the controller controlling the operation of the load assembly in an automated manner.

2. The apparatus of claim 1 wherein the sample of biomolecular analyte is disposed about a bead that is magnetically attractable, and wherein the load assembly includes an electromagnetic loading device that, in response to the controller, (i) electromagnetically carries the bead from a sample source to the test module using electromagnetism, and (ii) releases the bead into the test module.

3. The apparatus of claim 2 wherein the load assembly further includes an electromagnetic unloading device that provides a force on the bead in a direction toward the test module and away from the electromagnetic loading device in response to the controller.

4. The apparatus of claim 1 wherein the load assembly includes a capillary that, in response to the controller enabling supply of a voltage between the sample source and test module, (i) transfers the sample from a sample source to the test module using electrokinetics, and (ii) terminates transfer of the sample.

5. The apparatus of claim 4 wherein the load assembly further includes a gasket that forms a hermetic seal between the capillary and the test module when the capillary transfers the sample from the sample source to the test module.

6. The apparatus of claim 1 further comprising:

a detection assembly, coupled to the support assembly, that detects the components within the sample as separated by electrophoresis.

7. The apparatus of claim 6 wherein the detection assembly includes:

an actuating member coupled to the support assembly; and

a scanner, coupled to the actuating member, that scans the test module, the actuating member moving the scanner, in response to the controller, between (i) a first position adjacent the support assembly that receives and supports the test module and (ii) a second position adjacent the support assembly that receives and supports another test module.

8. The apparatus of claim 1 further comprising:

an injection assembly, coupled to the support assembly, that injects fluid serving as a matrix into the test module prior to loading the test module with the biomolecular analyte sample, such that the fluid facilitates separation of the components within the sample by electrophoresis.

9. The apparatus of claim 1 further comprising:

a set of electrical connections that provides power from an additional power supply to the test module.

10. The apparatus of claim 1 wherein the load assembly includes:

a robotic device, coupled to the support assembly, having an arm and an actuator that moves the arm between a sample source and the test module;

a loading device, coupled to the arm of the robotic device, that transfers the sample of biomolecular analyte from the sample source to the test module; and

a camera, coupled to the arm of the robotic device, that determines a position of the arm of the robotic device and indicates the position to the controller, the controller moving the robotic device according to the indicated position.

11. The apparatus of claim 1 wherein the support assembly includes a support member that supports multiple test modules and wherein the apparatus further comprises:

a detector that moves relative to the support member to scan each of the multiple test modules supported by the support member.

12. The apparatus of claim 1 wherein the support assembly includes a support member that supports multiple test modules, and wherein the apparatus further comprises:

a detector that scans the multiple test modules, the support member moving the multiple test modules relative to the detector such that the detector scans each test module.

13. A system for analyzing short tandem repeats within a sample of biomolecular analyte, comprising:

a test plate having a microfabricated separation channel optimized for high throughput processing;

a support assembly that supports the test plate;

an automated loading device, coupled to the support assembly, that loads the sample of biomolecular analyte on the test plate in an automated manner;

an electrophoresis power assembly, coupled to the support assembly, that effects electrophoretic separation of short tandem repeats in the sample within the microfabricated separation channel of the test plate, the microfabricated channel defining a micron amount of the sample such that short tandem repeats electrophoretically separate out of the sample at a high throughput rate; and

a high speed detector coupled to the support assembly for enabling counting, in a high throughput manner, of short tandem repeats electrophoretically separated out of the sample.

14. The system of claim 13, wherein the automated loading device includes a robotic actuator assembly, coupled to the support assembly, that obtains the sample from a sample source and deposits the sample at a particular location on the test plate.

15. The system of claim 13 wherein the automated loading device includes a capillary assembly, coupled to the support assembly, that obtains the sample from a sample source and deposits the sample at a particular location on the test plate.

16. A method for analyzing short tandem repeats within a sample of biomolecular analyte, comprising the steps of:

providing a test plate having a microfabricated separation channel optimized for high throughput;

providing a support assembly that supports the test plate;

activating an automated loading device that loads the sample of biomolecular analyte on the test plate in an automated manner;

connecting an electrophoresis power assembly, coupled to the support assembly to the test plate and activating the electrophoresis power assembly to effect separation of short tandem repeats in the sample within the microfabricated separation channel of the test plate, the microfabricated channel defining a micron amount of the sample such that short tandem repeats electrophoretically separate out of the sample at a high throughput rate; and

counting in a high throughput manner, the short tandem repeats electrophoretically separated out of the sample.

17. The method of claim 16 wherein the step of activating the automated loading device includes the step of:

enabling a robotic actuator assembly to obtain the sample from a sample source and deposit the sample at a particular location on the test plate.

18. The method of claim 16 wherein the step of activating the automated loading device includes the step of:

enabling a capillary assembly to obtain the sample from a sample source and deposit the sample at a particular location on the test plate.

19. The method of claim 16, wherein the step of counting includes

detecting the short tandem repeats within the separation channel of the test plate.