Separation of polynucleotides using supports having a plurality of electrode-containing cells

• US 5,434,049 A
• Filed: 02/24/1993
• Issued: 07/18/1995
• Est. Priority Date: 02/28/1992
• Status: Expired due to Fees

• Current Assignee: Hitachi, Ltd.
• Original Assignee: Hitachi, Ltd.
• Inventors: Kambara, Hideki, Okano, Kazunori
• Primary Examiner(s): Parr, Margaret
• Assistant Examiner(s): Horlick, Kenneth R.
• Application Number:
  US08/021,667
• Time in Patent Office: 874 Days
• Field of Search: 435/6, 436/94, 436/72, 436/807, 436/809, 536/25.4, 422/101, 422/68.1
• US Class Current: 435/6.11
• CPC Class Codes: B01J 2219/00653 the compounds being bound to electrodes embedded in or on the solid supports
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: Y10S 436/807 Apparatus included in process claim, e.g. physical support structures
• CPC Class Codes: Y10S 436/809 Multifield plates or multicontainer arrays
• CPC Class Codes: Y10T 436/143333 Saccharide [e.g., DNA, etc.]

Abstract

A method for simultaneously detecting a plurality of target polynucleotides in a sample on a single reaction chip and a method for separating a plurality of target polynucleotides are provided. On the reaction chip are arranged a plurality of independent cells for capturing different target polynucleotides. Different probes are immobilized onto the individual cells, and labeled probes detect the cells where the target polynucleotides are captured, thereby analyzing the species of the captured target polynucleotides. Also, the individual cells function as electrodes to elute the captured target polynucleotides therethrough, thereby separating a plurality of the target polynucleotides.

Claims

1. A method for separating polynucleotides in a sample solution comprising the steps of;

immobilizing individually different probes complementary to a plurality of target polynucleotides onto a plurality of cells formed at different regions on a single polynucleotide capturing support having an electrode on a surface of each of the cells,

adding the sample solution to the surface of the support and covering the surface of each cell with the sample solution, for capturing the target polynucleotides onto said polynucleotide capturing support by a hybridization reaction, and

supplying an electric field onto the electrode of each cell separately to elute the hybridized target polynucleotides from the cells individually.

2. A method according to claim 1, further comprising a step wherein a capillary provided with another electrode is placed upon each of said cells and the hybridized target polynucleotide on each of said cells is eluted and collected into the capillary individually.

3. A method for separating polynucleotides in a sample solution comprising;

immobilizing a plurality of different probes, complementary to a plurality of different target polynucleotides, onto surfaces of a plurality of cells, wherein each cell has only one probe immobilized on a cell surface, each cell is formed at a different region on a single polynucleotide capturing support, and each cell has an electrode on a cell surface,

adding the sample solution to the polynucleotide capturing support and covering each cell surface with the sample solution,

hybridizing the target polynucleotides to said probes immobilized on the polynucleotide capturing support under hybridization conditions that permit each probe to specifically hybridize to a complementary target polynucleotide,

supplying an electric field to each electrode of each cell separately to separately elute the hybridized target polynucleotides from the cells individually, and

recovering eluted target polynucleotides.

4. A method according to claim 3, further comprising

placing a capillary provided with an electrode upon each of said cells before supplying the electric field to each of said cells to elute hybridized target polynucleotide from each of said cells and

collecting an eluted target polynucleotide into the capillary.

5. A method according to claim 3, further comprising washing the cells with an aqueous solution to remove unhybridized polynucleotides before supplying an electric field.