Controlled electrophoresis method

• US 20070134812A1
• Filed: 11/17/2006
• Published: 06/14/2007
• Est. Priority Date: 11/07/2003
• Status: Active Grant

• Current Assignee: Princeton Biochemicals, Inc.
• Original Assignee: Princeton Biochemicals, Inc.
• Inventors: Guzman, Norberto
• Granted Patent: US 8,030,092 B2
• Time in Patent Office: Days
• US Class Current: 436/517
• 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 33/543 with an insoluble carrier for immobilising immunochemicals
• 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

1-321. -321. (canceled)

322. A controlled electrophoresis method, comprising:

introducing a sample into a transport passage of an electrophoresis apparatus;

the electrophoresis apparatus including a first separation passage intersecting the transport passage at a first overlapping portion, a first analyte concentrator at the first overlapping portion to concentrate a first analyte of interest, a second separation passage intersecting the transport passage at a second overlapping portion, and a second analyte concentrator at the second overlapping portion to concentrate a second analyte of interest;

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

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

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

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

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

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

delivering the released first analyte of interest in the first separation passage to a detector system which identifies and characterizes the first analyte of interest; and

delivering the released second analyte of interest in the second separation passage to the detector system which also identifies and characterizes the second analyte of interest.

323. The method of claim 322 wherein the localizing the first analyte concentrator is after the passing into the first overlapping portion and the localizing the second analyte concentrator is after the passing into the second overlapping portion.

324. The method of claim 322 wherein the localizing the first analyte concentrator includes closing valves associated with the first overlapping portion.

325. The method of claim 322 wherein the localizing the second analyte concentrator includes closing valves associated with the second overlapping portion.

326. The method of claim 322 wherein the localizing the first analyte concentrator includes heating same.

327. The method of claim 322 further comprising before the releasing the first analyte of interest, subjecting the first analyte concentrator to at least one of gentle shaking, microwave pulsing or acoustic mixing.

328. The method of claim 322 wherein the releasing the first analyte of interest includes introducing an eluting solution into the first separation passage upstream of the first analyte concentrator.

329. A controlled electrophoresis method, comprising:

introducing a sample into a transport passage;

passing the introduced sample past a first portion of the transport passage which intersects with an overlapping first separation passage so that a first analyte of interest in the sample is captured by a first analyte concentrator in the first portion;

controlling the temperature of the first analyte concentrator to a first temperature associated with the first analyte concentrator;

passing the introduced sample past a second portion of the transport passage which intersects with an overlapping second separation passage so that a second analyte of interest in the sample is captured by a second analyte concentrator in the second portion;

controlling the temperature of the second analyte concentrator to a different second temperature associated with the second analyte concentrator;

releasing the captured first analyte from the first analyte concentrator into the first separation passage;

causing the released first analyte to pass by electrophoresis migration or by a combination of electrophoresis migration and pressure to a detector system which identifies and characterizes the first analyte;

releasing the captured second analyte from the second analyte concentrator into the second separation passage; and

causing the released second analyte to pass by electrophoresis migration or by a combination of electrophoresis migration and pressure to the detector system which also identifies and characterizes the second analyte of interest.

330. The method of claim 329 wherein the controlling the temperature of the first analyte concentrator includes closing valves surrounding the first analyte concentrator.

331. The method of claim 329 wherein the first and second temperatures differ by about 12 degrees Centigrade.

332. The method of claim 329 wherein one of the first and second temperatures is approximately 25 degrees Centigrade and the other is approximately 37 degrees Centigrade.

333. The method of claim 329 further comprising before the releasing the captured first analyte, subjecting the first analyte concentrator to gentle shaking.

334. The method of claim 329 further comprising before the releasing the captured first analyte, subjecting the first analyte concentrator to microwave pulsing.

335. The method of claim 329 further comprising before the releasing the captured first analyte, subjecting the first analyte concentrator to acoustic mixing.

336. The method of claim 329 wherein the releasing the captured first analyte includes introducing an eluting solution into the first separation passage.

337. The method of claim 329 wherein the controlling the temperature of the first analyte concentrator includes isolating the first analyte concentrator by closing valves associated therewith.