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Related Concept Videos

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...
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...
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...
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.
In HPLC, two phases play a critical role in the separation process:
Size-Exclusion Chromatography01:08

Size-Exclusion Chromatography

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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Related Experiment Video

Updated: Jun 19, 2026

Separation of Single-stranded DNA, Double-stranded DNA and RNA from an Environmental Viral Community Using Hydroxyapatite Chromatography
13:46

Separation of Single-stranded DNA, Double-stranded DNA and RNA from an Environmental Viral Community Using Hydroxyapatite Chromatography

Published on: September 29, 2011

Protein chromatography on hydroxyapatite columns.

Larry J Cummings1, Mark A Snyder, Kimberly Brisack

  • 1Bio-Rad Laboratories, Inc., Hercules, California, USA.

Methods in Enzymology
|November 7, 2009
PubMed
Summary

Spherical hydroxyapatite chromatography enables efficient protein purification, reducing impurities like host cell proteins and DNA. This method enhances protein load capacity and column longevity for biopharmaceutical applications.

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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

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Last Updated: Jun 19, 2026

Separation of Single-stranded DNA, Double-stranded DNA and RNA from an Environmental Viral Community Using Hydroxyapatite Chromatography
13:46

Separation of Single-stranded DNA, Double-stranded DNA and RNA from an Environmental Viral Community Using Hydroxyapatite Chromatography

Published on: September 29, 2011

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

Area of Science:

  • Biochemistry
  • Chemical Engineering
  • Chromatography

Background:

  • Spherical hydroxyapatite offers advantages for protein scientists in purification.
  • Biopharmaceutical companies utilize single and multiple column applications for complex samples.

Purpose of the Study:

  • To review the use of spherical hydroxyapatite in protein purification.
  • To discuss adsorption/desorption mechanisms and column lifetime factors.
  • To cover column packing methods and diverse applications.

Main Methods:

  • Review of adsorption and desorption mechanisms of hydroxyapatite.
  • Analysis of chemical interactions affecting column lifetime.
  • Evaluation of column packing methods for spherical and microcrystalline hydroxyapatite.

Main Results:

  • Multiple column purification allows higher protein loads and better impurity reduction.
  • Hydroxyapatite interactions with ions, metals, and phosphates impact column longevity.
  • Low ionic strength buffers can cause significant pH shifts due to hydroxonium ion dynamics.

Conclusions:

  • Spherical hydroxyapatite is a versatile tool for protein purification, particularly for recombinant proteins and monoclonal antibodies.
  • Understanding hydroxyapatite's multimodal properties is key to optimizing purification strategies.
  • Robust packing methods and buffer selection are crucial for maximizing column performance and lifespan.