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Related Concept Videos

Chromatography: Introduction01:10

Chromatography: Introduction

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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...
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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.
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Analyte Adsorption and Distribution01:09

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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...
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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.
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Principles Of Column Chromatography01:13

Principles Of Column Chromatography

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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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Ion-Exchange Chromatography01:09

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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...
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A direct comparison between membrane adsorber and packed column chromatography performance.

Cristiana Boi1, Andrea Malavasi2, Ruben G Carbonell3

  • 1DICAM, Alma Mater Studiorum-Università di Bologna, Bologna, Italy.

Journal of Chromatography. A
|November 1, 2019
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Summary

This study compared packed bed and membrane chromatography for protein purification. Membrane chromatography achieved higher productivity at high flow rates, despite lower binding capacities than packed beds.

Keywords:
Binding capacityChromatographyMembrane adsorberPacked columnProductivity

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Area of Science:

  • Biotechnology
  • Chemical Engineering
  • Separation Science

Background:

  • Chromatography is crucial for protein purification.
  • Packed bed and membrane chromatography are common methods.
  • Optimizing these methods is key for efficient bioprocessing.

Purpose of the Study:

  • To compare the performance of packed bed and membrane chromatography for protein purification.
  • To evaluate anion exchange chromatography using a strong Q ligand and bovine serum albumin (BSA) as a model protein.
  • To assess equilibrium and dynamic binding capacities, productivity, and bed utilization.

Main Methods:

  • Utilized anion exchange media with a strong Q quaternary ammonium group.
  • Employed bovine serum albumin (BSA) as the model protein.
  • Maintained constant stationary phase volume (3 mL) and mobile phase superficial velocity for both packed bed and membrane adsorbers.

Main Results:

  • Packed bed columns exhibited higher equilibrium binding (66.9 mg/mL) and dynamic binding capacities (62.8 mg/mL at 97.5 cm/h) compared to membrane adsorbers (43.04 mg/mL and 20.7 mg/mL, respectively).
  • Membrane adsorbers achieved a maximum productivity 3.3 times higher (111 mg/(mL h)) than packed columns.
  • Bed utilization was higher in packed beds at long residence times but favored membrane chromatography at short residence times.

Conclusions:

  • Packed beds offer higher binding capacities due to greater surface area.
  • Membrane chromatography demonstrates superior performance and productivity at high flow rates.
  • The choice between packed bed and membrane chromatography depends on desired flow rates and productivity targets in protein purification.