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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...
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...
Types Of Column Chromatography01:29

Types Of Column Chromatography

The stability and compatibility of column material with samples are crucial for efficient purification in chromatographic techniques. Various operating parameters such as pH, temperature, or solvent affect the packing of the column material, thereby determining the purification efficiency. The choice of column material also plays an essential role in deciding the operating parameters and can be modified based on the proteins that need to be purified.
Gel Filtration Chromatography
When the...
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...
Thin-Layer Chromatography (TLC): Overview01:11

Thin-Layer Chromatography (TLC): Overview

Thin-layer chromatography (TLC) is a chromatography technique that separates compounds based on their polarity. TLC typically uses polar silica gel, a form of silicon dioxide, as the stationary phase. The silica gel contains hydroxyl (OH) groups on its surface, which form hydrogen bonds with polar compounds, influencing their adhesion to the stationary phase.
To begin the analysis, a mixture of compounds is spotted on the starting line on the TLC plate using a thin capillary. The bottom of the...
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...

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Erratum to: 10th asilomat conference on mass spectrometry: Time of flight mass spectrometry.

Journal of the American Society for Mass Spectrometry·2013
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Immobilized metal ion affinity chromatography.

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Direct evidence of the generation in human stomach of an antimicrobial peptide domain (lactoferricin) from ingested lactoferrin.

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Direct detection and quantitative determination of bovine lactoferricin and lactoferrin fragments in human gastric contents by affinity mass spectrometry.

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Related Experiment Video

Updated: Jun 3, 2026

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
10:21

Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification

Published on: September 21, 2011

Thiophilic adsorption chromatography.

T W Hutchens1

  • 1Department of Pediatrics, Baylor College of Medicine, Houston, TX.

Methods in Molecular Biology (Clifton, N.J.)
|March 25, 2011
PubMed
Summary

Thiophilic adsorption efficiently purifies immunoglobulins under mild conditions. This method uniquely binds three major immunoglobulin classes and selectively removes them from biological fluids for various applications.

Area of Science:

  • Biochemistry
  • Protein Chemistry
  • Affinity Chromatography

Background:

  • Immunoglobulin purification is crucial for research and therapeutics.
  • Existing methods often require harsh conditions or lack broad applicability.
  • Thiophilic adsorption offers a novel affinity purification approach.

Purpose of the Study:

  • To highlight the utility of thiophilic adsorption for immunoglobulin purification.
  • To demonstrate its unique capacity for adsorbing multiple immunoglobulin classes.
  • To showcase its application in selective immunoglobulin depletion from biological fluids.

Main Methods:

  • Utilizing thiophilic adsorption as an affinity matrix for immunoglobulin binding.
  • Employing mild, neutral pH conditions for immunoglobulin recovery.

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Thin-layer Chromatographic (TLC) Separations and Bioassays of Plant Extracts to Identify Antimicrobial Compounds

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Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
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  • Applying the method for selective depletion in complex biological samples.

    • Main Results:

    • Thiophilic adsorption effectively purifies three major immunoglobulin classes and subclasses.
    • Immunoglobulins are recovered efficiently at neutral pH without structural damage.
    • Selective depletion of immunoglobulins from serum, hybridoma media, and milk is achieved.

    Conclusions:

    • Thiophilic adsorption is a versatile and mild technique for immunoglobulin purification.
    • Its unique binding capacity and efficient recovery make it valuable for research.
    • Applications include hybridoma technology and infant nutrition studies.