Two-dimensional gels for separation, identification and characterization of biomolecules

• US 6,507,664 B1
• Filed: 08/17/2000
• Issued: 01/14/2003
• Est. Priority Date: 06/24/1997
• Status: Expired due to Fees

• Current Assignee: Large Scale Biology Corporation
• Original Assignee: Large Scale Biology Corporation
• Inventors: Goodman, Jack, Anderson, N. Leigh, Anderson, Norman G.
• Primary Examiner(s): Johns, Andrew W.
• Application Number:
  US09/642,246
• Time in Patent Office: 880 Days
• Field of Search: 382/128, 382/129, 382/153, 702/19, 204/461, 204/607, 204/612, 436/516, 435/7.1, 435/288.7, 700/266, 700/273
• US Class Current: 382/129
• CPC Class Codes: G01N 2001/288 Filter punches
• CPC Class Codes: G01N 27/44773 Multi-stage electrophoresis, e.g. two-dimensional electrophoresis
• CPC Class Codes: G01N 27/44782 of a plurality of samples
• CPC Class Codes: G01N 27/44795 Isoelectric focusing
• CPC Class Codes: Y10T 436/113332 with conveyance of sample along a test line in a container or rack
• CPC Class Codes: Y10T 436/25 including sample preparation
• CPC Class Codes: Y10T 436/25375 Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]
• CPC Class Codes: Y10T 436/255 including use of a solid sorbent, semipermeable membrane, or liquid extraction

Abstract

The present invention provides an integrated, fully automated, high-throughput system for two-dimensional electrophoresis comprised of gel-making machines, gel processing machines, gel compositions and geometries, gel handling systems, sample preparation systems, software and methods. The system is capable of continuous operation at high-throughput to allow construction of large quantitative data sets.

Claims

1. A computer-assisted method for isolating one or more selected proteins present in a two-dimensional array prepared by two-dimensional electrophoresis in a loose polyacrylamide gel, the method comprising:

(a) imaging the two-dimension array or a replica thereof with a first device to generate a computer-readable output;

(b) processing the output in at least one computer to select a protein for isolation;

(c) generating machine-readable instructions that direct a second device to isolate the selected protein from the two-dimensional array; and

(d) isolating the selected protein with the second device in accordance with the machine-readable instructions by removing a portion of the loose polyacrylamide gel containing the selected protein, wherein the second device is physically separate from the first device.

2. A method for studying proteins separated into an array contained in a loose gel, the method comprising:

(a) imaging the array of proteins with a first device;

(b) selecting one or more members of the array for removal, and

(c) removing those portions of the loose gel that contain the selected members of the array by means of a second device under remote computer control, the second device comprising an excision component capable of performing x and y movements, wherein the second device is physically separate from the first device.

3. A computer-readable medium comprising instructions for causing a computer to perform the following steps:

selecting a protein for isolation by processing output generated by a first device upon imaging a two-dimensional gray of proteins present in a loose polyacrylamide gel; and

generating machine-readable instructions that direct a second device to isolate the selected protein from the two-dimensional array by removing a portion of the loose polyacrylamide gel containing the selected protein, wherein the second device is physically separate from the first device.

4. A computer comprising a program for performing the following steps:

selecting a protein for isolation by processing output generated by a first device upon imaging a two-dimensional array of proteins present in a loose polyacrylamide gel; and

generating machine-readable instructions that direct a second device to isolate the selected protein from the two-dimensional ay by removing a portion of the loose polyacrylamide gel contain the selected protein, when the second device is physically separate from the first device.