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Related Concept Videos

Electrophoresis: Overview01:20

Electrophoresis: Overview

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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...
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Capillary Electrophoresis: Applications01:30

Capillary Electrophoresis: Applications

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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,...
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Capillary Electrophoresis: Instrumentation01:20

Capillary Electrophoresis: Instrumentation

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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...
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Two-dimensional Gel Electrophoresis01:22

Two-dimensional Gel Electrophoresis

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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...
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Size-Exclusion Chromatography01:08

Size-Exclusion Chromatography

2.4K
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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SDS-PAGE01:27

SDS-PAGE

35.3K
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...
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Related Experiment Video

Updated: Mar 16, 2026

Assembly and Characterization of Polyelectrolyte Complex Micelles
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Using capillary electrophoresis to characterize polymeric particles.

Kathryn R Riley1, Sophia Liu2, Guo Yu3

  • 1Department of Chemistry, Wake Forest University, Winston-Salem, NC 27109, USA; Material Measurement Laboratory, National Institute of Standards and Technology, Gaithersburg, MD 20899, USA.

Journal of Chromatography. A
|August 21, 2016
PubMed
Summary
This summary is machine-generated.

Capillary electrophoresis (CE) effectively characterizes polymeric particles by size and surface properties. Binding studies reveal aptamer-conjugated particles exhibit a 5-fold higher dissociation constant compared to free aptamers.

Keywords:
Binding constantsCapillary electrophoresisDNA-conjugated particlesIon sphere particlesNECEEMPolystyrene particles

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Area of Science:

  • Analytical Chemistry
  • Materials Science
  • Biotechnology

Background:

  • Polymeric micro- and sub-micron particles are crucial in various applications, requiring precise characterization.
  • Understanding particle properties like size, surface functionality, and binding affinity is essential for optimizing their use.
  • Capillary electrophoresis (CE) offers a powerful technique for high-resolution separation and analysis of such particles.

Purpose of the Study:

  • To develop and validate a capillary zone electrophoresis (CZE) method for characterizing diverse polymeric particles.
  • To demonstrate the separation of specific particle types, including DNA-templated particles.
  • To investigate the impact of particle conjugation on molecular binding properties using an aptamer-protein system.

Main Methods:

  • Development of a robust capillary zone electrophoresis (CZE) method for particle separation and quantitation.
  • Characterization of polystyrene particles with varying surface modifications (amino, carboxylate, sulfate) and sizes (0.2-3.0 μm).
  • Demonstration of separating DNA-templated polyacrylamide particles from untemplated ones.
  • Determination of dissociation constants (Kd) for aptamer-protein interactions in free solution and on particle surfaces.

Main Results:

  • Baseline separation and quantitation of various commercially available polystyrene particles were achieved using CZE.
  • Successful differentiation between DNA-templated and untemplated polyacrylamide particles was demonstrated.
  • Dissociation constants for the thrombin aptamer and thrombin protein were determined.
  • A significant increase (approximately 5-fold) in dissociation constant was observed for the aptamer when conjugated to a particle compared to its free solution state.

Conclusions:

  • Capillary electrophoresis is a versatile and effective technique for the comprehensive characterization of polymeric micro- and sub-micron particles.
  • Particle surface functionality and size significantly influence their separation and behavior in CE.
  • Conjugating aptamers to particles can alter their binding affinity, with implications for selection and binding processes in solid-phase applications.
  • The findings provide valuable insights into particle characterization and the behavior of biomolecular interactions on solid supports.