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Facile method and apparatus for the analysis of biological macromolecules in two dimensions using common and familiar electrophoresis formats

a technology of biological macromolecules and electrophoresis, which is applied in the direction of fluid pressure measurement, liquid/fluent solid measurement, peptide measurement, etc., can solve the problems of difficult two-dimensional separation of complex mixtures using polyacrylamide electrophoresis, and achieves easy two-dimensional gel electrophoresis, reduces streaking, and reduces the time without electric power

Inactive Publication Date: 2009-09-17
LIFE TECH CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The patent describes a method for pre-casting and partially dehydrating isoelectric focusing gels onto a semi-rigid support, and a manifold for absorbing and rehydrating the gels. The gels are then flushed with water and mounted into a vertical electroporesis apparatus for further separation. The method allows for the simultaneous loading of samples and the rapid diffusion of reagents for the second dimension separation. The patent also provides pre-cast second dimension gradient gels and a surface mounted horizontal opening or port for sealing the isoelectric focusing strip to the second dimension gel. The technical effects of the patent include improved efficiency and accuracy in isoelectric focusing and electrophoresis separation.

Problems solved by technology

This two-dimensional separation of complex mixtures using polyacrylamide electrophoresis is a powerful but difficult technique.

Method used

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  • Facile method and apparatus for the analysis of biological macromolecules in two dimensions using common and familiar electrophoresis formats
  • Facile method and apparatus for the analysis of biological macromolecules in two dimensions using common and familiar electrophoresis formats
  • Facile method and apparatus for the analysis of biological macromolecules in two dimensions using common and familiar electrophoresis formats

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example 1

Step 1: Mount Dehydrated Strips on Manifold

[0057]FIG. 5A shows partially dehydrated pre-cast polyacrylamide pH gradient gel strips (1), bound to a perforated plastic backing (2) at 9.0 millimeter spacing, are pressed into molded channels (8) on one side of an elastomeric manifold (6) so that each strip is sealed with a laminae space above the gel. Each laminar space is accessible for filling through two normally closed diagonal ports, top and bottom on the opposite side of the elastomeric manifold from the strips.

Step 2: Load Samples

[0058]FIG. 5B shows how the elastomeric manifold (6) is mounted at an angle on the diagonal support stand (7). Samples containing proteins to be focused are loaded through the bottom ports (10), pushing open the collapsible channels (11) and filling the laminae space above each strip. Sample loading is most easily accomplished using an 8-place pipettor. Sets of disposable pipette tips are first inserted in the top ports. The tips hold the top port (10) o...

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Abstract

An ensemble of components and methods are disclosed for utilizing two-dimensional electrophoresis in polyacrylamide or related polymer gels using common, existing and familiar electrophoresis formats and equipment. The disclosed invention makes two-dimensional electrophoresis convenient and easy to use for individuals already using vertical “mini-gel” type systems. The invention discloses the combination of pre-cast disposable gels for both first and second separation dimensions using novel support devices and cassettes that simplify the difficult multiple sample handling and processing steps inherent in ordinary two-dimensional electrophoresis methods. Devices and methods are disclosed in said invention that provide exceptional gel to gel reproducibility.

Description

[0001]This application claims priority from U.S. Provisional Application No. 60 / 147,490 filed on Aug. 9, 1999.TECHNICAL FIELD AND INDUSTRIAL APPLICABILITY OF INVENTION[0002]The invention relates to the production of acrylamide or other gels for use in separations of proteins, nucleic acids or other biological materials. The invention further relates to making two-dimensional electrophoresis convenient and easy to use for individuals already using vertical “mini-gel” type systems. The invention discloses the combination of pre-cast disposable gels for both first and second separation dimensions using novel support devices and cassettes that simplify the difficult multiple sample handling and processing steps inherent in ordinary two-dimensional electrophoresis methods.BACKGROUND OF THE INVENTION[0003]Polyacrylamide gel electrophoresis has become one of the most frequently used techniques for the separation of biological macromolecules such as proteins, nucleic acids and polysaccharid...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): G01N27/26B01D57/02
CPCG01N27/44795G01N27/44782
Inventor CHAMPAGNE, JAMES T.
Owner LIFE TECH CORP
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