Related Experiment Video
Updated: Aug 3, 2026
10:21
Automated Hydrophobic Interaction Chromatography Column Selection for Use in Protein Purification
Published on: September 21, 2011
Large scale protein separations: engineering aspects of chromatography
1Industrial Biotechnology Centre, University of Waterloo, Ontario, Canada.
Biotechnology Advances
|January 1, 1990
Summary
This review covers large-scale protein purification using industrial chromatography, focusing on ion exchange and size exclusion techniques. Advances in biopolymer separation will enhance biotechnology production.
Area of Science:
- Biotechnology
- Chemical Engineering
- Biochemistry
Background:
- Industrial-scale chromatographic purification is crucial for producing biopharmaceuticals.
- Understanding scale-up challenges is essential for efficient protein separation.
Purpose of the Study:
- To review engineering considerations for large-scale protein purification.
- To discuss the impact of operational parameters and column characteristics on separation performance.
Main Methods:
- Review of industrial chromatography fundamentals.
- Analysis of separation column geometry, operational parameters, and packing materials.
- Emphasis on ion exchange and size exclusion chromatography.
Main Results:
- Key factors affecting large-scale separation include column geometry, operational parameters, and packing characteristics.
- Ion exchange and size exclusion chromatography are dominant techniques in commercial protein purification.
- Current technology is assessed, identifying areas for future development.
Conclusions:
- Physico-chemical advances will significantly improve chromatographic separation of biopolymers.
- Chromatography will play an increasingly vital role in the industrial production of novel biotechnology products.
Related Concept Videos
01:13Principles 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...
01:22Two-dimensional Gel Electrophoresis
Two-dimensional gel electrophoresis is a high-resolution protein separation method first introduced by O' Farrell and Klose in 1975. This method involves protein separation by two dimensions, mass and charge, making it more accurate than one-dimensional gel electrophoresis.
The first dimension separation uses the isoelectric focusing or IEF technique performed on immobilized pH gradient (IPG) strips that separate proteins according to their isoelectric points.
Biological samples, such as cells...
The first dimension separation uses the isoelectric focusing or IEF technique performed on immobilized pH gradient (IPG) strips that separate proteins according to their isoelectric points.
Biological samples, such as cells...
01:29Types 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...
Gel Filtration Chromatography
When the...
01:15Optimizing Chromatographic Separations
Optimizing chromatographic separations is crucial for obtaining clean separations in a minimum amount of time. Optimization is required for several factors, including kinetic effects related to band broadening, plate height, capacity factor, and separation factor.
Band broadening refers to spreading solute bands as they travel through the column. This broadening can impact resolution. Plate height (H) represents the length required for one theoretical plate. A lower plate height corresponds to...
Band broadening refers to spreading solute bands as they travel through the column. This broadening can impact resolution. Plate height (H) represents the length required for one theoretical plate. A lower plate height corresponds to...
01:09Ion-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...
01:08Size-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,...
Silica particles offer advantages such as rigidity,...