Microfabricated capillary electrophoresis chip and method for simultaneously detecting multiple redox labels
First Claim
1. A microfabricated capillary electrophoresis chip including a substrate with an elongated, narrow separation channel which widens into a detection reservoir and means for applying a separation voltage along the channel, two or more thin film working electrodes each adapted to receive a different working voltage extending into said detection reservoir opposite and near the end of said narrow separation channel where the working electrodes have minimal influence from the high electrophoresis potentials, said working electrodes simultaneously detecting current generated by molecules undergoing redox reaction as they migrate past the thin film electrodes after they have migrated down the channel and a ground electrode adapted to be connected to said means for applying a separation voltage along the channel and a reference electrode adapted to be connected to a reference voltage, both in said detection reservoir spaced from said working electrodes.
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Abstract
This invention relates to a microfabricated capillary electrophoresis chip for detecting multiple redox-active labels simultaneously using a matrix coding scheme and to a method of selectively labeling analytes for simultaneous electrochemical detection of multiple label-analyte conjugates after electrophoretic or chromatographic separation.
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Citations
46 Claims
- 1. A microfabricated capillary electrophoresis chip including a substrate with an elongated, narrow separation channel which widens into a detection reservoir and means for applying a separation voltage along the channel, two or more thin film working electrodes each adapted to receive a different working voltage extending into said detection reservoir opposite and near the end of said narrow separation channel where the working electrodes have minimal influence from the high electrophoresis potentials, said working electrodes simultaneously detecting current generated by molecules undergoing redox reaction as they migrate past the thin film electrodes after they have migrated down the channel and a ground electrode adapted to be connected to said means for applying a separation voltage along the channel and a reference electrode adapted to be connected to a reference voltage, both in said detection reservoir spaced from said working electrodes.
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8. A method of selective electrochemical detection of analytes in a complex mixture of analytes which comprises the steps of:
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labeling each analyte in the mixture with a redox label which generates an electrochemical signal which is different from the label attached to other analytes, separating the mixture to provide individual labeled analytes, and simultaneously detecting with two or more working electrodes the different electrochemical signals generated by the redox labels on the separated analytes to identify the individual analytes. - View Dependent Claims (9, 10, 11, 12, 13, 14)
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15. The method of selective electrochemical detection of analytes in a complex mixture of analytes which comprises the steps of:
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labeling each analyte in the mixture with a redox label which generates an electrochemical signal which is different from the label attached to other analytes, separating the mixture to provide individual labeled analytes, and simultaneously detecting the different electrochemical signals generated by the redox labels on the separated analytes to identify the individual analytes. - View Dependent Claims (16, 17, 18, 19, 20, 21)
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22. A method of electrochemically detecting individual analytes in a mixture of analytes comprising the steps of:
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labeling each analyte in the mixture with a redox label which generates an electrochemical signal which is different from the label attached to other analytes, separating the mixture in an electrophoretic separation channel, placing two or more electrochemical detection electrodes at or near the end of said separation channel, and simultaneously detecting the different electrochemical signals generated by the redox labels at each of said one or more detection electrodes to uniquely identify the individual labeled analytes. - View Dependent Claims (23, 24, 25)
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26. A method of determining the sequence of a DNA template which comprises the steps of:
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generating and redox labeling all possible complementary sequencing fragments of the DNA template to be sequenced where the sets of fragments terminating with the four different bases (A, C, G, T) are identified by distinct electrochemical signals generated by the redox label associated with each of the distinct sets or fragments, electrophoretically separating said sets of labeled fragments in a single channel or lane, and simultaneously detecting the distinct electrochemical signals generated by the redox labels to identify the individual fragments. - View Dependent Claims (27, 28, 29, 30, 31, 32, 33, 34)
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35. A method of performing nucleic acid genotyping using electrochemical detection comprising the steps of:
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labeling each nucleic acid analyte in a mixture with a unique redox active label which generates an electrochemical signal which is different from the labels attached to the other analytes, electrophoretically separating the mixture in a gel-filled capillary or channel, placing two or more electrochemical detection electrodes at or near the end of said separation capillary or channel, and simultaneously detecting the different electrochemical signals generated by the redox labels at each of said detection electrodes to uniquely identify the individual labeled analytes. - View Dependent Claims (36, 37, 38, 39, 40, 41, 42)
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43. A method of selective electrochemical detection of analytes in a complex mixture of intrinsically redox active analytes which comprises the steps of:
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electrophoretically separating the mixture in a capillary or channel, placing two or more electrochemical detection electrodes at or near the end of said separation capillary or channel, and simultaneously detecting the different electrochemical signals generated by the redox active analytes at each of said detection electrodes to uniquely identify the individual analytes. - View Dependent Claims (44, 45, 46)
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Specification