Capillary electrophoresis apparatus and method

• US 5,560,811 A
• Filed: 03/21/1995
• Issued: 10/01/1996
• Est. Priority Date: 03/21/1995
• Status: Expired due to Fees

• Current Assignee: Soane Biosciences, Inc.
• Original Assignee: Seurat Analytical Systems, Inc.
• Inventors: Hoyt, David W., Briggs, Jonathan, McCormick, Randy M.
• Primary Examiner(s): Niebling, John
• Assistant Examiner(s): Wong, Edna
• Application Number:
  US08/408,683
• Time in Patent Office: 560 Days
• Field of Search: 204/299 R, 204/180.1
• US Class Current: 204/451
• CPC Class Codes: B01L 9/065 specially adapted for capillary tubes
• CPC Class Codes: G01N 2035/00237 Handling microquantities of analyte, e.g. microvalves, capillary networks
• CPC Class Codes: G01N 27/44743 Introducing samples
• CPC Class Codes: G01N 27/44782 of a plurality of samples

Abstract

This invention involves method and apparatus for multiplexing electrophoresis analysis. An array of samples in multi well plates are simultaneously transferred to an array of electrophoresis column where electrophoresis is simultaneously carried out followed by analysis of the columns. The methods and apparatus of this invention are, for example, useful for DNA analysis, including sequencing, and for measuring reactions between specifically binding proteins and their binding partners.

Claims

1. An electrophoresis separation plate comprising:

a) a transportable frame having first and second ends and having an array of electrophoresis capillaries with first and second ends mounted respectively between the first and second ends of the frame;

b) the first end of the frame having a buffer reservoir and an electrode in the buffer reservoir wherein the first end of the capillaries are in liquid communication with the buffer reservoir; and

c) the second end of the frame having a means for placing the second end of the capillaries in contact with an array of liquid samples or run buffer which is in contact with electrodes and wherein there is fluid communication between the buffer reservoir and the array of samples or the run buffer through the electrophoresis capillaries and electrical communication between the electrode in the buffer reservoir and the electrodes in the samples or run buffer by way of the electrophoresis capillaries.

2. A system for capillary electrophoresis analysis of an array of samples in an array of sample containers comprising:

a) means for simultaneously transferring at least a portion of each sample in the array of samples to a corresponding array of capillary electrophoresis columns wherein the capillary electrophoresis columns are in the electrophoresis plate of claim 1;

b) means for simultaneously conducting capillary electrophoresis separations on the array of transferred samples in the capillary electrophoresis columns; and

c) means for analyzing capillary electrophoresis separations from (b).

5. A method for analysis of an array of samples in an array of sample containers by capillary electrophoresis comprising:

a) providing an array of samples in an array of sample wells;

b) simultaneously transferring at least a portion of each sample in the array of sample wells to a corresponding array of capillary electrophoresis columns wherein the capillary electrophoresis columns are in the electrophoresis separation plate of claim 1;

c) simultaneously conducting separation of the transferred samples by capillary electrophoresis; and

d) analyzing the capillary electrophoresis separations of (c).

7. A system according to claim 2 wherein the array of samples conforms to the format of a 96, 192 or 384 well plate.

8. A separation plate according to claim 1 wherein the plate contains 8, 12, 16 and 24 electrophoresis capillaries spaced apart to match the spacing in a row or columns of 8, 12, 16 or 24 wells in a 96, 192, or 384 well plate.

11. The method of claim 5 wherein the array of samples is nucleic acid of varying length.

12. The method of claim 5 wherein the array of samples is a specifically binding protein, its binding partner or the complex of the two.

15. The electrophoresis separation plate of claim 1 wherein there is provided adjacent to sample handling plate wells a buffer wash well and a run buffer well.

16. The electrophoresis separation plate of claim 1 wherein electrodes are mounted in the frame near the second end of the capillaries so that when the capillaries are in the samples or the run buffer there is electrical communication between the electrode in the buffer reservoir and the electrodes near the capillaries.

3. An apparatus for processing an array of samples from an array of sample wells comprising:

a) a sample handling plate which defines an array of sample handling plate wells wherein each well has a sipper capillary in liquid communication with the sample handling plate well to provide an array of sipper capillaries and wherein a porous matrix is interposed between each sipper capillary and each sample handling plate well, wherein the array of sipper capillaries will simultaneously wick an array of samples from an array of sample wells;

b) a base plate which defines an opening to receive the sample handling plate and defines an inner chamber to means associated with the base plate and sample handling plate to seal the inner chamber and provide a sealed inner chamber; and

c) means for pressurizing and evacuating the sealed inner chamber to move liquid on either side of the porous matrix to the other side of the porous matrix.

9. The apparatus of claim 3 wherein the sipper capillaries of the sample handling plate wells are in an array which conforms to a format of a 96, 192 or 384 well plate.

10. The apparatus of claim 3 wherein the means for pressurizing and evacuating is programmed to alternately pressurize and evacuate the inner chamber to mix reagents added above the porous matrix with sample below the porous matrix.

17. The apparatus of claim 3 wherein the porous matrix is a membrane.

4. Electrophoresis separation plate comprising:

a frame having a buffer reservoir at one end and a plurality of sample sites at the other end and having an electrode at each end,

a means for mounting capillary electrophoresis columns on the frame so that there is electrical communication between the electrodes and fluid communication between the sample sites and reservoir when there is fluid in the sample sites, capillary electrophoresis columns and buffer reservoir.

6. A method for analysis of an array of samples in an array of sample containers by capillary electrophoresis comprising:

a) providing the array of samples in an array of sample wells;

b) simultaneously acquiring an aliquot of each sample from the array of samples with a sample handling plate having an array of sample handling plate wells with sipper capillaries;

c) transferring the sample handling plate to a base plate which provides for simultaneously processing and then presenting the samples for capillary electrophoresis;

d) simultaneously transferring at least a portion of the presented samples to an array of capillary electrophoresis columns;

e) simultaneously conducting separations by capillary electrophoresis of the presented transferred samples; and

f) analyzing the capillary electrophoresis separation of e.

13. The method of claim 6 wherein one or more reagents are added to the sample handling plate wells and mixed with the samples.

14. The method of claim 6 wherein the array of electrophoresis columns are rinsed for reuse to separate additional samples.