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Related Concept Videos

Affinity Chromatography01:03

Affinity Chromatography

Affinity chromatography is a powerful technique extensively utilized for separating and purifying specific biomolecules from complex mixtures. It capitalizes on the highly selective binding between an analyte and its counterpart, such as antibody-antigen interactions. The counterpart is immobilized on the stationary phase, forming an affinity column. The stationary phase typically consists of solid support, such as agarose or porous glass beads, immobilizing the affinity ligand. The mobile...
Analyte Adsorption and Distribution01:09

Analyte Adsorption and Distribution

In certain chromatographic separations, solutes transfer between the mobile phase and the stationary phase via sorption, which typically refers to the process of adsorption. For many chromatographic systems, the sorption process often depends on the polarity of the compounds—an expression of the overall dipole moment within the molecule. During the separation process, there is competition between the solute and solvent for adsorption to the stationary phase. Highly polar compounds and solvents...
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...
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...
Adsorption Isotherms II01:25

Adsorption Isotherms II

Brunauer, Emmett, and Teller (BET) introduced a theory in 1938 that modified Langmuir's assumptions to explain multilayer physical adsorption. This theory is applicable to Type II isotherms and provides a more realistic picture of adsorption processes. The BET theory assumes a uniform solid surface with localized adsorption sites, where adsorption at one site doesn't affect adsorption at neighboring sites. This theory also allows for the possibility of additional molecules being adsorbed on top...
Gas Chromatography: Types of Columns and Stationary Phases01:17

Gas Chromatography: Types of Columns and Stationary Phases

Gas chromatography (GC) relies on stationary phases to separate and analyze components in a sample. There are two main types of stationary phases: liquid and solid. Liquid stationary phases are non-volatile, thermally stable, and chemically inert liquids coated onto the column. Solid stationary phases are particles of adsorbent material, such as silica gel or molecular sieves.
For an analyte to remain on the column for a sufficient amount of time, it must exhibit some level of compatibility (or...

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Related Experiment Video

Updated: Jul 5, 2026

Activated Cross-linked Agarose for the Rapid Development of Affinity Chromatography Resins - Antibody Capture as a Case Study
07:53

Activated Cross-linked Agarose for the Rapid Development of Affinity Chromatography Resins - Antibody Capture as a Case Study

Published on: August 16, 2019

Expanded-bed adsorption chromatography.

Robert M Kennedy1

  • 1GE Healthcare, Piscataway, New Jersey, USA.

Current Protocols in Protein Science
|April 23, 2008
PubMed
Summary
This summary is machine-generated.

Expanded-bed adsorption (EBA) chromatography enables direct protein capture from unclarified samples. This technique efficiently separates target proteins from contaminants using flow manipulation for elution.

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Last Updated: Jul 5, 2026

Activated Cross-linked Agarose for the Rapid Development of Affinity Chromatography Resins - Antibody Capture as a Case Study
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Area of Science:

  • Biotechnology
  • Chromatographic techniques
  • Protein purification

Background:

  • Traditional chromatography requires pre-clarified samples, adding processing steps.
  • Unclarified samples contain cells and debris that can clog conventional columns.
  • Expanded-bed adsorption (EBA) chromatography offers a solution for direct processing.

Purpose of the Study:

  • To describe the principles and application of expanded-bed adsorption chromatography.
  • To highlight EBA's utility in capturing proteins directly from crude biological mixtures.
  • To outline the operational modes and post-elution procedures in EBA.

Main Methods:

  • Expansion of the adsorbent bed using upward buffer flow.
  • Loading of unclarified feed through the expanded bed, capturing target proteins.
  • Elution of captured proteins using a change in buffer composition, with options for expanded-bed or settled-bed elution.

Main Results:

  • EBA effectively captures target proteins while allowing particulates and contaminants to pass through.
  • Both expanded-bed and settled-bed elution modes are demonstrated, adaptable to feed characteristics.
  • Post-elution cleaning-in-place (CIP) and regeneration procedures ensure column longevity.

Conclusions:

  • Expanded-bed adsorption chromatography is a versatile and efficient method for direct protein capture from crude samples.
  • The technique simplifies downstream processing by eliminating the need for sample clarification.
  • EBA facilitates robust protein purification workflows with options for elution and column maintenance.