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Related Concept Videos

Capillary Electrophoresis: Applications01:30

Capillary Electrophoresis: Applications

Capillary electrophoretic separations offer various modes, each with unique applications. These modes include capillary zone electrophoresis, capillary gel electrophoresis, capillary array electrophoresis, capillary isoelectric focusing, capillary isotachophoresis, micellar electrokinetic chromatography, and capillary electrochromatography.
Capillary zone electrophoresis (CZE) separates ionic components based on their electrophoretic mobility. It has been used to separate proteins, amino acids,...
Electrophoresis: Overview01:20

Electrophoresis: Overview

Electrophoresis is a powerful analytical separation technique that relies on the differential migration of charged species when subjected to an electric field. The core strength of electrophoresis lies in its ability to separate high-molecular-weight species in complex mixtures. It has found widespread use in biochemistry, molecular biology, and analytical chemistry, allowing the separation of compounds like amino acids, nucleotides, carbohydrates, and proteins with excellent resolution.
There...
Capillary Electrophoresis: Instrumentation01:20

Capillary Electrophoresis: Instrumentation

Capillary electrophoresis instrumentation typically consists of several key components. A high-voltage power supply generates the electric field necessary for the separation by connecting to an anode (the positively charged electrode) and a cathode (the negatively charged electrode) located in buffer reservoirs at each end of the capillary tube. The system includes a sample vial, a fused silica capillary tube coated with polyimide for mechanical strength through which the sample components...
SDS-PAGE01:27

SDS-PAGE

Gel electrophoresis is a method that separates biological macromolecules like nucleic acids or proteins by forcing them to pass through a gel matrix under an electric field.
A variation of gel electrophoresis, termed  polyacrylamide gel electrophoresis (PAGE), is commonly used for separating proteins according to their molecular size by passing them through a polyacrylamide gel. Because of the varying charges associated with amino acid side chains, PAGE can be used to separate intact proteins...
Two-dimensional Gel Electrophoresis01:22

Two-dimensional Gel Electrophoresis

Two-dimensional gel electrophoresis is a high-resolution protein separation method first introduced by O' Farrell and Klose in 1975. This method involves protein separation by two dimensions, mass and charge, making it more accurate than one-dimensional gel electrophoresis.
The first dimension separation uses the isoelectric focusing or IEF technique performed on immobilized pH gradient (IPG) strips that separate proteins according to their isoelectric points.
Biological samples, such as  cells...
Size-Exclusion Chromatography01:08

Size-Exclusion Chromatography

In size-exclusion chromatography (SEC), also known as molecular-exclusion or gel-permeation chromatography, molecules are separated based on their sizes. This technique is important for separating large molecules such as polymers and biomolecules. The two classes of micron-sized stationary phases encountered in SEC are silica particles and cross-linked polymer resin beads. Both materials are porous, but their pore sizes vary significantly.
Silica particles offer advantages such as rigidity,...

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Capillary Electrophoresis to Monitor Peptide Grafting onto Chitosan Films in Real Time
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Published on: October 26, 2016

Transformable capillary electrophoresis for oligosaccharide separations using phospholipid additives.

Ruijuan Luo1, Stephanie A Archer-Hartmann, Lisa A Holland

  • 1C. Eugene Bennett Department of Chemistry, West Virginia University, Morgantown, West Virginia 26506, USA.

Analytical Chemistry
|January 19, 2010
PubMed
Summary
This summary is machine-generated.

Phospholipids enhance glycan separation in capillary electrophoresis by optimizing composition, hydration, and temperature. This method improves resolution and expands applications for analyzing complex carbohydrate structures.

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Area of Science:

  • Analytical Chemistry
  • Biochemistry
  • Separation Science

Background:

  • Glycan analysis is crucial for understanding biological processes.
  • Capillary electrophoresis (CE) is a powerful separation technique.
  • Phospholipids can be utilized as additives to improve CE performance.

Purpose of the Study:

  • To investigate the use of phospholipids as additives in CE for enhanced glycan separation.
  • To determine the optimal conditions for phospholipid-enhanced CE separation.
  • To explore the integration of lectins and enzymes for expanded glycan analysis.

Main Methods:

  • Capillary electrophoresis was employed to separate glycans.
  • Phospholipid preparations with varying compositions (DMPC/DHPC ratios), hydration, and temperatures were tested.
  • Separation performance was evaluated based on efficiency and resolution.
  • Concanavalin A and alpha1-2,3 mannosidase were incorporated into the separation system.

Main Results:

  • A specific phospholipid preparation (1,2-dimyristoyl-sn-glycero-3-phosphocholine and 1,2-dihexanoyl-sn-glycero-3-phosphocholine at a 2.5 ratio) in a 10% lipid/aqueous buffer at 25°C yielded optimal separation efficiency at 400 V/cm.
  • The addition of phospholipids significantly enhanced resolution.
  • Incorporation of Concanavalin A and alpha1-2,3 mannosidase provided additional glycan selectivity.

Conclusions:

  • Phospholipid additives can significantly improve the separation of glycans in capillary electrophoresis.
  • Optimizing phospholipid properties and CE conditions is key to achieving high separation efficiency and resolution.
  • The combined use of phospholipid additives with lectins and enzymes offers a versatile approach for advanced glycan analysis.