Electrophoresis standards, methods and kits

• US 8,945,361 B2
• Filed: 09/20/2006
• Issued: 02/03/2015
• Est. Priority Date: 09/20/2005
• Status: Active Grant

• Current Assignee: ProteinSimple (Bio-Techne Corporation)
• Original Assignee: ProteinSimple (Bio-Techne Corporation)
• Inventors: Gentalen, Erik, Suich, Daniel J.
• Primary Examiner(s): Van, Luan
• Assistant Examiner(s): Kaur, Gurpreet
• Application Number:
  US11/524,630
• Publication Number:
  US 20070062813A1
• Time in Patent Office: 3,058 Days
• Field of Search: 204451-455, 204/548, 204601-605, 204/644
• US Class Current: 204/548
• CPC Class Codes: C07K 1/13 Labelling of peptides
• CPC Class Codes: C07K 1/26 Electrophoresis
• CPC Class Codes: C07K 1/28 Isoelectric focusing
• CPC Class Codes: C07K 7/06 having 5 to 11 amino acids
• CPC Class Codes: C07K 7/08 having 12 to 20 amino acids gastrins C07K14/595; somatostatins C07K14/655; melanotropins C07K14/68
• CPC Class Codes: G01N 2550/00 Electrophoretic profiling, e.g. for proteome analysis
• CPC Class Codes: G01N 27/44726 using specific dyes, markers or binding molecules

Abstract

Electrophoresis Compositions, methods and kits useful for, among other things, detecting, quantifying and/or characterizing analytes are provided. The compositions are useful as electrophoresis standards for determine the isoelectric point and molecular weight of an analyte. The electrophoresis standards generally comprise at least one label moiety and one or more reactive moieties that when activated attach the standard to a substrate.

Claims

1. A method of determining the isoelectric point of one or more analytes by isoelectric focusing comprising the steps of:

forming a pH gradient in a fluid path including one or more analytes and one or more standards, the one or more standards including one or more moieties, at least one of the one or more moieties being a at least one reactive moiety, when the at least one reactive moiety is activated to form an activated moiety, the activated moiety attaches the one or more standards to a substrate;

focusing the one or more analytes at a position equal to the isoelectric point of each analyte from the one or more analytes and focusing the one or more standards at a position equal to the isoelectric point of each standard from the one or more standards;

immobilizing the one or more analytes and the one or more standards in said fluid path by activating the at least one reactive moiety, the immobilizing being non-specific to any analyte from the one or more analytes;

detecting one or more signals generated from the one or more analytes and the one or more standards; and

determining the isoelectric point of each analyte from the one or more analytes by comparing the signals of the one or more analytes to the signals of the one or more standards.

2. The method of claim 1, wherein said fluid path is comprised of one or more capillaries.

3. The method of claim 1, wherein said fluid path is comprised of one or more channels in a microfluidic device.

4. The method of claim 1, wherein said fluid path is comprised of one or more gels.

5. A method of determining the isoelectric point of one or more analytes by isoelectric focusing comprising the steps of:

generating an electric field to form a pH gradient in a fluid path including one or more analytes and one or more standards, the one or more standards including one or more reactive moieties configured to attach the one or more standards to a substrate;

immobilizing the one or more analytes and the one or more standards in said fluid path by activating the one or more reactive moieties, the immobilizing being non-specific to any analyte from the one or more analytes;

detecting one or more signals generated from the one or more analytes and the one or more standards when the one or more analytes and the one or more standards are at their respective isoelectric point; and

generating data associated with a determination of the isoelectric point of each analyte from the one or more analytes by comparing the signals of the one or more analytes to the signals of the one or more standards.

6. The method of claim 5, wherein the one or more reactive moieties are activated to form at least one activated moiety, the activated moiety configured to attach the one or more standards to the substrate.

7. The method of claim 5, further comprising:

before the detecting, focusing the one or more analytes at a position equal to the isoelectric point of each analyte from the one or more analytes and focusing the one or more standards at a position equal to the isoelectric point of each standard from the one or more standards.

8. The method of claim 5, wherein the fluid path includes one of one or more capillaries, a microfluidic device or one or more gels.