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Related Concept Videos

Principles Of Column Chromatography01:13

Principles Of Column Chromatography

The chromatography technique was first invented in 1901 by Michael S. Tswett, a Russian botanist, to separate plant pigments using organic solvents. Further, in 1941, Archer John Porter Martin and R. L. M. Synge modified the technique by packing silica gel into a column. A mixture of amino acids was then separated on the packed column using chloroform and water mixture as the mobile phase. This was the first report on column chromatography. At present, column chromatography is a widely used...
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Diffusion on Chromatography Columns

In column chromatography, when an analyte is introduced as a narrow band at the top of the column, the solutes begin to separate and broaden, developing a Gaussian profile. This broadening occurs due to various factors, such as longitudinal diffusion.
Longitudinal diffusion occurs when the solute molecules in the mobile phase diffuse from the more concentrated center of the chromatographic band to the more dilute regions on either side, both towards and against the flow direction. This...
Chromatography: Introduction01:10

Chromatography: Introduction

Chromatography is a technique used to separate compounds based on differences of partitioning between two phases, the stationary phase and the mobile phase.
The phase in which the compounds linger or on which the compounds adsorb is called the stationary phase, whereas the mobile phase is the solvent that carries the solutes to be analyzed. In traditional column chromatography, the mixture flows through the stationary phase, and the compounds partition between the stationary and mobile phases...
High-Performance Liquid Chromatography: Elution Process01:05

High-Performance Liquid Chromatography: Elution Process

In High-Performance Liquid Chromatography (HPLC), the elution process is critical to the separation of analytes and the quality of chromatographic results. Elution describes how compounds move through the column and separate based on their interactions with the mobile and stationary phases. This process determines the resolution, peak shape, and retention times in the chromatogram, which are essential for identifying and quantifying components in complex mixtures. Understanding the elution...
Ion-Exchange Chromatography01:09

Ion-Exchange Chromatography

Ion-exchange chromatography, or IEC, is a technique for separating ions based on their affinity for the stationary phase. The stationary phase is a cross-linked polymer resin with covalently attached ionic functional groups. The functional groups can be either positively charged (cation exchangers) or negatively charged (anion exchangers). A cation exchanger consists of a polymeric anion and active cations, while an anion exchanger is a polymeric cation with active anions. The choice of...
High-Performance Liquid Chromatography: Introduction01:11

High-Performance Liquid Chromatography: Introduction

High-performance liquid chromatography(HPLC), formerly referred to as High-pressure liquid chromatography, is a powerful technique used to separate, identify, and quantify components in complex mixtures. The term "high pressure" refers to using high pressure to push the liquid mobile phase through the tightly packed columns.
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Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
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Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification

Published on: September 21, 2011

Countercurrent gradient chromatography: A continuous focusing technique.

L L Evans1, M A Burns

  • 1Department of Chemical Engineering, University of Michigan, Ann Arbor, Michigan 48109-2136.

Biotechnology and Bioengineering
|December 5, 1995
PubMed
Summary
This summary is machine-generated.

This study introduces a magnetically stabilized fluidized bed for continuous protein separation using countercurrent gradient chromatography (CGC). The system achieves stable pH gradients for efficient protein purification and concentration.

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Chromatographic Fingerprinting by Template Matching for Data Collected by Comprehensive Two-Dimensional Gas Chromatography
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Area of Science:

  • Biochemistry
  • Chromatography
  • Chemical Engineering

Background:

  • Continuous separation techniques are crucial for protein purification.
  • Existing methods may lack efficiency or stability in gradient formation.
  • Countercurrent chromatography offers potential for improved separation.

Purpose of the Study:

  • To develop and validate a continuous protein separation system using countercurrent gradient chromatography (CGC).
  • To investigate the use of a magnetically stabilized fluidized bed (MSFB) for establishing stable pH gradients.
  • To model and optimize the CGC system for protein concentration and purity.

Main Methods:

  • Utilized a magnetically stabilized fluidized bed (MSFB) to create a countercurrent flow between a solid resin and a liquid buffer.
  • Established stable pH gradients within the CGC system in under 10 minutes.
  • Separated myoglobin and human serum albumin (HSA) to assess system performance.
  • Developed a parameter-free mathematical model to predict system behavior.

Main Results:

  • Achieved stable pH gradients for over 2 hours.
  • Demonstrated controllable gradient shapes by adjusting phase velocities.
  • Obtained concentration factors of 1.75 for myoglobin and 1.2 for HSA.
  • The mathematical model accurately predicted system focusing behavior.

Conclusions:

  • The MSFB-based CGC system provides a stable and controllable platform for continuous protein separation.
  • The developed mathematical model aids in optimizing parameters for enhanced purity and concentration.
  • This technology offers a promising approach for efficient protein purification and concentration.