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Electrophoresis: Overview

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: Instrumentation

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...

Two-dimensional Gel Electrophoresis

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...

Capillary Electrophoresis: Applications

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,...

SDS-PAGE

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...

Chromatographic Resolution

Chromatographic Resolution

In chromatography, a solute moves through a chromatographic column and tends to spread, forming a Gaussian-shaped band. The longer the solute spends in the column, the broader the band becomes. The broadening can lead to overlaps within the column, affecting separation effectiveness.
The effectiveness of separation can be evaluated by determining the level of separation between two neighboring peaks in a chromatogram, which represents the individual components of a sample.
In chromatography,...

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Related Experiment Video

Updated: Jun 12, 2026

The Fabrication and Operation of a Continuous Flow, Micro-Electroporation System with Permeabilization Detection

The Fabrication and Operation of a Continuous Flow, Micro-Electroporation System with Permeabilization Detection

Published on: January 7, 2022

High resolution continuous flow electrophoresis.

W A Gobie¹, J B Beckwith, C F Ivory

¹Department of Chemical Engineering University of Notre Dame Notre Dame, Indiana 46556.

Biotechnology Progress

|June 23, 2010

Summary

This summary is machine-generated.

Continuous free-flow electrophoresis (CFE) can be enhanced by recycling effluent, overcoming limitations like diffusion. This recycling method allows for high-purity recovery of biomolecules, improving preparative-scale purification processes.

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Area of Science:

Biochemistry
Analytical Chemistry
Chemical Engineering

Background:

Continuous free-flow electrophoresis (CFE) is a preparative technique for purifying biomolecules.
Classic CFE suffers from limited solute resolution due to diffusion and dispersion effects.
These limitations hinder the purification of sensitive biomolecules at scale.

Purpose of the Study:

To address the limitations of classic CFE for biomolecule purification.
To introduce a modified CFE technique with effluent recycling.
To develop mathematical models for predicting separation performance in the modified system.

Main Methods:

Derivation of mathematical models for a recycled continuous flow electrophoresis (RCFE) device.
Analysis of two limiting cases: rapid diffusion and slow diffusion.
Utilizing analytical solutions to predict separation factors under varied operational conditions.

Main Results:

The RCFE technique effectively overcomes diffusion and dispersion limitations inherent in classic CFE.
Complete recovery of undiluted product is achievable.
Arbitrarily high purities can be obtained through continuous recycling.

Conclusions:

Recycling effluent in CFE significantly enhances solute resolution and purification efficiency.
The developed mathematical models accurately predict separation performance for RCFE.
RCFE offers a superior method for preparative-scale purification of labile biomolecules.