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US 6,387,234 B1Granted
Integrated multiplexed capillary electrophoresis system
AssigneeIowa State University Research Foundation, Inc.
InventorsEdward S. Yeung, Hongdong Tan
Life of this patent
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Abstract
The present invention provides an integrated multiplexed capillary electrophoresis system for the analysis of sample analytes. The system integrates and automates multiple components, such as chromatographic columns and separation capillaries, and further provides a detector for the detection of analytes eluting from the separation capillaries. The system employs multiplexed freeze/thaw valves to manage fluid flow and sample movement. The system is computer controlled and is capable of processing samples through reaction, purification, denaturation, pre-concentration, injection, separation and detection in parallel fashion. Methods employing the system of the invention are also provided.
Claims46
An integrated multiplexed capillary electrophoresis system for analysis of an analyte in a sample comprising:
- a plurality of intake capillaries;
- a plurality of first junctions, each first junction in fluid communication with one of the plurality of intake capillaries;
- a first manifold in fluid communication with the first junctions;
- a chromatographic column array comprising a plurality of chromatographic columns having an outlet end, each chromatographic column in fluid communication with one of the plurality of first junctions;
- a first detector positioned to detect analytes eluting from the outlet ends of the chromatographic columns to determine whether the analyte arrival times are synchronous;
- a plurality of second junctions, each second junction in fluid communication with one of the plurality of chromatographic columns, the chromatographic columns being interposed between the first and second junctions;
- a second manifold in fluid communication with the second junctions;
- and
- a separation capillary array comprising a plurality of separation capillaries, each separation capillary in fluid communication with one of the plurality of second junctions and having an outlet end;
- the system having a first configuration wherein the chromatographic columns are in fluid communication with the intake capillaries and the separation capillaries and a second configuration wherein the chromatographic columns are in fluid communication with the first and second manifolds.
7 more elements
A method for detecting an analyte in a plurality of samples comprising:
- (a) providing an integrated multiplexed capillary electrophoresis system comprising
- a plurality of intake capillaries, each intake capillary having an intake end;
- a plurality of first junctions, each first junction in fluid communication with one of the plurality of intake capillaries;
- a first manifold in fluid communication with the first junctions;
- a chromatographic column array comprising a plurality of chromatographic columns having an outlet end, each chromatographic column in fluid communication with one of the plurality of first junctions;
- a plurality of second junctions, each second junction in fluid communication with one of the plurality of chromatographic columns, the chromatographic columns being interposed between the first and second junctions;
- a second manifold in fluid communication with the second junctions;
- and
- a separation capillary array comprising a plurality of separation capillaries, each separation capillary in fluid communication with one of the plurality of second junctions and having an outlet end;
- the system having a first configuration wherein the chromatographic columns are in fluid communication with the intake capillaries and the separation capillaries and a second configuration wherein the chromatographic columns are in fluid communication with the first and second manifold;
- (b) introducing each of the plurality of samples into the inlet end of a different intake capillary;
- (c) transferring each of the plurality of samples from each intake capillary into a chromatographic column in fluid communication with each of the intake capillaries;
- (d) chromatographing each of the plurality of samples to yield a plurality of purified sample portions, the purified sample portions comprising at least one detectable analyte;
- (e) injecting each of the plurality of purified sample portions into a separation capillary in fluid communication with the chromatographic column;
- (f) separating each of the purified sample portions to yield a plurality of separated sample portions, each of the separated sample portions comprising at least one detectable analyte;
- and
- (g) detecting at least one detectable analyte.
14 more elements
A method for sequencing nucleic acids in a plurality of samples comprising:
- (a) providing an integrated multiplexed capillary electrophoresis system comprising
- a plurality of intake capillaries at least one intake capillary comprising a reaction portion for reacting a sample;
- a plurality of first junctions, each first junction in fluid communication with one of the plurality of intake capillaries;
- a first manifold in fluid communication with the first junctions;
- a chromatographic column array comprising a plurality of chromatographic columns having an outlet end, each chromatographic column in fluid communication with one of the plurality of first junctions;
- a plurality of second junctions, each second junction in fluid communication with one of the plurality of chromatographic columns, the chromatographic columns being interposed between the first and second junctions;
- a second manifold in fluid communication with the second junction;
- a separation capillary array comprising a plurality of separation capillaries, each separation capillary in fluid communication with one of the plurality of second junctions and having an outlet end;
- a plurality of first freeze-thaw valves for regulating the flow of fluids in the system, each first freeze-thaw valve comprising a portion of an intake capillary wherein each of the plurality of first freeze-thaw valves comprises a distal freeze thaw valve comprising a portion of the intake capillary distal to the reaction portion and a proximal freeze-thaw valve comprising a portion of the intake capillary proximal to the reaction portion;
- a plurality of first joining capillaries, each first joining capillary interposed between a first junction and a chromatographic column, the first junction being in fluid communication with the intake capillary;
- a plurality of second freeze-thaw valves for regulating the flow of fluids in the system, each second freeze-thaw comprising a portion of a first joining capillary;
- (b) introducing each of the plurality of samples into the reaction portion of a different intake capillary, each sample comprising a nucleic acid;
- (c) reacting each of the plurality of samples in a DNA sequencing reaction to yield a plurality of reacted samples, each reacted sample comprising a plurality of detectably labeled nucleic acid fragments having different lengths;
- (d) transferring each of the plurality of reacted samples from each of the intake capillaries into the chromatographic column in fluid communication with the intake capillary;
- (e) chromatographing each of the plurality of reacted samples to yield a plurality of purified samples, each purified sample comprising a plurality of detectably labeled nucleic acid fragments having different lengths;
- (f) injecting each of the plurality of purified samples into a separation capillary in fluid communication with the chromatographic column;
- (g) separating each of the plurality of purified samples to yield, for each purified sample, a plurality of separated detectably labeled nucleic acid fragments having different lengths;
- (h) detecting the detectably nucleic acid fragments having different lengths;
- and
- (i) for each sample, determining the sequence of the nucleic acid.
17 more elements
Dependencies are inferred from the claim text.