Electrophoresis process using a valve system

• US 8,182,746 B2
• Filed: 10/31/2007
• Issued: 05/22/2012
• Est. Priority Date: 11/07/2003
• Status: Active Grant

• Current Assignee: Princeton Biochemicals, Inc.
• Original Assignee: Princeton Biochemicals, Inc.
• Inventors: Guzman, Norberto A.
• Primary Examiner(s): Jung, Unsu
• Application Number:
  US11/981,479
• Publication Number:
  US 20080223722A1
• Time in Patent Office: 1,665 Days
• Field of Search: None
• US Class Current: 422/82.01
• CPC Class Codes: G01N 1/405 by adsorption or absorption
• CPC Class Codes: G01N 27/44708 Cooling
• CPC Class Codes: G01N 27/44743 Introducing samples
• CPC Class Codes: G01N 27/44756 Apparatus specially adapted therefor
• CPC Class Codes: G01N 33/561 Immunoelectrophoresis

Abstract

An electrophoresis apparatus is generally disclosed for sequentially analyzing a single sample or multiple samples having one or more analytes in high or low concentrations. The apparatus comprises a relatively large-bore transport capillary which intersects with a plurality of small-bore separation capillaries and includes a valve system. Analyte concentrators, having antibody-specific (or related affinity) chemistries, are stationed at the respective intersections of the transport capillary and separation capillaries to bind one or more analytes of interest. The apparatus allows the performance of two or more dimensions for the optimal separation of analytes. The apparatus may also include a plurality of valves surrounding each of the analyte concentrators to localize each of the concentrators to improve the binding of one or more analytes of interest.

Claims

1. A controlled electrophoresis method, comprising:

introducing a sample into a transport passage of a multi-dimensional electrophoresis apparatus;

the multi-dimensional electrophoresis apparatus including a first separation passage intersecting the transport passage at a first overlapping portion, a first biomarker concentrator at the first overlapping portion to concentrate a first biomarker of interest,

a second separation passage intersecting the transport passage at a second overlapping portion,

a second biomarker concentrator at the second overlapping portion to concentrate a second biomarker of interest,

the transport passage being communicable at an inlet end with a sample supply,

the first and second separation passages being communicable upstream of the first and second overlapping portions separate and independently with a biomarker separation buffer supply or with a supply of an elution buffer or solution,

a valve system for controlling the flow of the sample in the transport passage from the inlet end of the transport passage to an outlet end of the transport passage,

the valve system controlling the flow of the biomarker separation buffer in the separation passage introduced from a biomarker separation buffer supply or a plug of a biomarker elution buffer or solution in the separation passage introduced from a biomarker elution buffer supply in a sequential order from an inlet end of the separation passage to an outlet end of the separation passage,

the valve system comprising a first valve on the first separation passage downstream of the first biomarker concentrator,

a second valve on the transport passage upstream of the first biomarker concentrator,

a third valve on the transport passage downstream of the first biomarker concentrator,

a fourth valve on the first separation passage upstream of the first biomarker concentrator,

a fifth valve on the second separation passage upstream of the second biomarker concentrator,

a sixth valve on the second separation passage downstream of the second biomarker concentrator, and

a seventh valve on the transport passage downstream of the second biomarker concentrator,

passing the introduced sample in the transport passage into the first overlapping portion so that the first biomarker of interest in the sample is concentrated in the first biomarker concentrator;

passing the introduced sample in the transport passage into the second overlapping portion so that the second biomarker of interest in the sample is concentrated in the second biomarker concentrator;

localizing the first biomarker concentrator and thereby increasing the concentration of the first biomarker of interest by the first biomarker concentrator;

localizing the second biomarker concentrator and thereby increasing the concentration of the second biomarker of interest by the second biomarker concentrator;

releasing the first biomarker of interest concentrated in the first biomarker concentrator into the first separation passage;

releasing the second biomarker of interest concentrated in the second biomarker concentrator into the second separation passage;

delivering the released first biomarker of interest in the first separation passage to a biomarker detection zone which identifies and characterizes the first biomarker of interest; and

delivering the released second biomarker of interest in the second separation passage to the biomarker detection zone which also identifies and characterizes the second biomarker of interest;

the first overlapping portion has a staggered configuration which includes a first elongated concentration area in which the first biomarker concentrator is positioned; and

/or

the second overlapping has a staggered configuration which includes a second elongated concentration area in which the second biomarker concentrator is positioned.

2. The electrophoresis process of claim 1 wherein:

the localizing the first biomarker concentrator includes closing the first valve, the second valve, the third valve and the fourth valve; and

the localizing the second biomarker concentrator includes closing the third valve, the fifth valve, the sixth valve and the seventh valve.

3. The electrophoresis process of claim 1 wherein the localizing includes holding the sample solution in the localized first and/or second biomarker concentrators for a period of time to increase respective biomarker binding conditions therein.

4. The electrophoresis process of claim 1 wherein the second, third and seventh valves located in the transport passage are opened to introduce sample in the transport passage, wherein the third valve when open opens the transport passage between the first and second biomarker concentrators and which when closed closes the transport passage between the first and second biomarker concentrators.

5. The electrophoresis process of claim 1 further comprising:

introducing a buffer solution via a first auxiliary buffer passage into the first separation passage at a location that is spaced a distance downstream of the first biomarker concentrator and a distance upstream of the biomarker detection zone, wherein the introducing the buffer solution into the first separation passage includes operating one or more auxiliary valves of the multi-dimensional electrophoresis apparatus so that buffer solution from the first auxiliary buffer passage does not interact with the first biomarker concentrator; and

/or

introducing a buffer solution via a second auxiliary buffer passage into the second separation passage at a location that is spaced a distance downstream of the second biomarker concentrator and a distance upstream of the biomarker detection zone, wherein the introducing the buffer solution into the second separation passage includes operating one or more auxiliary valves of the multi-dimensional electrophoresis apparatus so that buffer solution from the second auxiliary buffer passage does not interact with the second biomarker concentrator.

6. The electrophoresis process of claim 1 wherein the biomarkers are small molecular mass substances, biomolecules and/or cells, subcellular organelles, and/or their respective modified and/or altered corresponding counterpart.

7. The electrophoresis process of claim 1 further comprising operating the valve system via a central processing unit (CPU) of the multi-dimensional electrophoresis apparatus to separately, independently and sequentially control functions of the first and second biomarker concentrators.

8. The electrophoresis process of claim 1 wherein:

the multi-dimensional electrophoresis apparatus includes a plurality of separations passages which includes the first and second separation passages;

the multi-dimensional electrophoresis apparatus includes a plurality of biomarker concentrators which includes the first and second biomarker concentrators; and

a plurality of different biomarkers are associated with at least one predetermined disease; and

further comprising;

detecting data relative to biomarkers released from at least some of the biomarker concentrators by the biomarker detection zone.

9. The electrophoresis process of claim 8 further comprising:

sending data associated with the detected data over the Internet to an evaluator; and

after the sending, receiving feedback over the Internet from the evaluator relative to the at least one predetermined disease.

10. The electrophoresis process of claim 9 wherein the evaluator is a medical specialist positioned remote from the electrophoresis apparatus.

11. The electrophoresis process of claim 8 further comprising:

using a central processing unit (CPU), for determining from the detected biomarker data a diagnosis of whether an individual has and/or is predisposed to having the at least one predetermined disease.

12. The electrophoresis process of claim 11 further comprising:

after determining, sending data associated with if the individual has and/or is predisposed to having the at least one predetermined disease and data associated with the detected biomarkers over the Internet from the central processing unit (CPU) to an evaluator and evaluating from the data associated with if the individual has and/or is predisposed to having the at least one predetermined disease and data associated with the detected biomarkers to provide biomarker evaluation feedback; and

the evaluator sending the biomarker evaluation feedback over the Internet to the central processing unit (CPU).

13. The electrophoresis process of claim 11 further comprising:

the central processing unit (CPU) comparing the biomarker evaluation feedback with reference data in a memory; and

updating the reference data in the memory based on the biomarker evaluation feedback.

14. The electrophoresis process of claim 8 wherein:

the at least one predetermined disease is one or more of diabetes, heart disease, cancer or other inflammatory disease; and

the specimen is urine, blood, hair, nail, sweat, cerebro-spinal fluid or other biological fluid or tissue or cell extract of the individual.

15. The electrophoresis process of claim 8 wherein:

the plurality of separation passages defines a first set of separation passages and a second set of separation passages; and

the plurality of biomarker concentrators defines a first set of biomarker concentrators associated with a first predetermined disease and with the first set of separation passages, and a second set of biomarker concentrators associated with a second predetermined disease and with the second set of separation passages.

16. The electrophoresis process of claim 15 wherein:

the first set of separation passages and the first set of biomarker concentrators form a first modular unit; and

the second set of separation passages and the second set of biomarker concentrators form an alternative second modular unit, the modular units may be connected to each other in a linear tandem fashion.

17. The electrophoresis process of claim 1 wherein the localizing the at least one of the first and second biomarker concentrators increases the concentration of the respective biomarker therein.

18. The electrophoresis process of claim 1 wherein the localizing the at least one of the first and second biomarker concentrators includes heating or cooling same.

19. The method of claim 1 wherein the delivering the released first biomarker of interest and the delivering the released second biomarker of interest includes the transfer of the one or more released first biomarker of interest bound to the first biomarker concentrator to the first separation passage, the transfer of the one or more released second biomarker of interest bound to the second biomarker concentrator to the second separation passage and the separation of the one or more released first biomarker of interest and one or more released second biomarker of interest respectively within the first and second separation passages independently, separate and sequentially, by one or more of any of the modes of capillary electrophoresis to the biomarker detection zone which detect, quantify, identify and characterizes the first or second biomarkers of interest by one or more detectors, independent, separate and in a sequential protocol.

20. A controlled electrophoresis method, comprising:

introducing a sample into a transport passage of a multi-dimensional electrophoresis apparatus;

the multi-dimensional electrophoresis apparatus including a first separation passage intersecting the transport passage at a first overlapping portion, a first biomarker concentrator at the first overlapping portion to concentrate a first biomarker of interest,

a second separation passage intersecting the transport passage at a second overlapping portion,

a second biomarker concentrator at the second overlapping portion to concentrate a second biomarker of interest,

the transport passage being communicable at an inlet end with a sample supply,

the first and second separation passages being communicable upstream of the first and second overlapping portions separate and independently with a biomarker separation buffer supply or with a supply of an elution buffer or solution,

a valve system for controlling the flow of the sample in the transport passage from the inlet end of the transport passage to an outlet end of the transport passage,

the valve system controlling the flow of the biomarker separation buffer in the separation passage introduced from a biomarker separation buffer supply or a plug of a biomarker elution buffer or solution in the separation passage introduced from a biomarker elution buffer supply in a sequential order from an inlet end of the separation passage to an outlet end of the separation passage,

the valve system comprising a first valve on the first separation passage downstream of the first biomarker concentrator,

a second valve on the transport passage upstream of the first biomarker concentrator,

a third valve on the transport passage downstream of the first biomarker concentrator,

a fourth valve on the first separation passage upstream of the first biomarker concentrator,

a fifth valve on the second separation passage upstream of the second biomarker concentrator,

a sixth valve on the second separation passage downstream of the second biomarker concentrator, and

a seventh valve on the transport passage downstream of the second biomarker concentrator,

passing the introduced sample in the transport passage into the first overlapping portion so that the first biomarker of interest in the sample is concentrated in the first biomarker concentrator;

passing the introduced sample in the transport passage into the second overlapping portion so that the second biomarker of interest in the sample is concentrated in the second biomarker concentrator;

localizing the first biomarker concentrator and thereby increasing the concentration of the first biomarker of interest by the first biomarker concentrator;

localizing the second biomarker concentrator and thereby increasing the concentration of the second biomarker of interest by the second biomarker concentrator;

releasing the first biomarker of interest concentrated in the first biomarker concentrator into the first separation passage;

releasing the second biomarker of interest concentrated in the second biomarker concentrator into the second separation passage;

delivering the released first biomarker of interest in the first separation passage to a biomarker detection zone which identifies and characterizes the first biomarker of interest; and

delivering the released second biomarker of interest in the second separation passage to the biomarker detection zone which also identifies and characterizes the second biomarker of interest;

the first set of separation passages and the first set of biomarker concentrators form a first modular unit; and

the second set of separation passages and the second set of biomarker concentrators form an alternative second modular unit;

before the introducing the sample solution, selecting one of the first and second modular units and operatively installing the selected one of the first and second modular units; and

removing the installed first or second modular unit and installing the other of the first and second modular units.

21. The method of claim 20 wherein the delivering the released first biomarker of interest and the delivering the released second biomarker of interest includes the transfer of the one or more released first biomarker of interest bound to the first biomarker concentrator to the first separation passage, the transfer of the one or more released second biomarker of interest bound to the second biomarker concentrator to the second separation passage and the separation of the one or more released first biomarker of interest and one or more released second biomarker of interest respectively within the first and second separation passages independently, separate and sequentially, by one or more of any of the modes of capillary electrophoresis to the biomarker detection zone which detect, quantify, identify and characterizes the first or second biomarkers of interest by one or more detectors, independent, separate and in a sequential protocol.