Expanded bed adsorption
Encyclopedia
Expanded bed adsorption (EBA) is a preparative chromatographic technique which makes processing of viscous and particulate liquids possible.
binding principles in EBA are the same as in classical column chromatography
and the common ion-exchange, hydrophobic interaction and affinity chromatography
ligands can be used. After the adsorption step is complete, the fluidized bed is washed to flush out any remaining particulates. Elution of the adsorbed proteins is commonly performed with the eluent flow in the reverse direction, that is, as a conventional packed bed, in order to recover the adsorbed solutes
in a smaller volume of eluent.
EBA may be considered to combine both the "Removal of Insolubles" and the "Isolation" steps of the 4-step downstream processing
heuristic. The major limitations associated with EBA technology is biomass interactions and aggregations onto adsorbent during processing.
Where classical column chromatography
uses a solid phase made by a packed bed, EBA uses particles in a fluidized
state. Expanded bed adsorption is, however, different from fluidised bed chormatography in essentially two ways: one, the EBA resin contains particles of varying size and density which results in a gradient of particle size when expanded; and two, when the bed is in its expanded state, local loops are formed. Particles such as whole cells or cell debris, which would clog a packed bed column, readily pass through a fluidized bed. EBA can therefore be used on crude culture broths or slurries of broken cells, thereby bypassing initial clearing steps such as centrifugation
and filtration
, which is mandatory when packed beds are used. The feed flow rate is kept low enough that the solid packing remains stratified and does not fluidize completely. Hence EBA can be modeled as frontal adsorption
in a packed bed, rather than as a well mixed continuous-flow adsorber.
Principle
The proteinProtein
Proteins are biochemical compounds consisting of one or more polypeptides typically folded into a globular or fibrous form, facilitating a biological function. A polypeptide is a single linear polymer chain of amino acids bonded together by peptide bonds between the carboxyl and amino groups of...
binding principles in EBA are the same as in classical column chromatography
Column chromatography
Column chromatography in chemistry is a method used to purify individual chemical compounds from mixtures of compounds. It is often used for preparative applications on scales from micrograms up to kilograms.The main advantage of column chromatography is the relatively low cost and disposability...
and the common ion-exchange, hydrophobic interaction and affinity chromatography
Affinity chromatography
Affinity chromatography is a method of separating biochemical mixtures and based on a highly specific interaction such as that between antigen and antibody, enzyme and substrate, or receptor and ligand.-Uses:Affinity chromatography can be used to:...
ligands can be used. After the adsorption step is complete, the fluidized bed is washed to flush out any remaining particulates. Elution of the adsorbed proteins is commonly performed with the eluent flow in the reverse direction, that is, as a conventional packed bed, in order to recover the adsorbed solutes
Solution
In chemistry, a solution is a homogeneous mixture composed of only one phase. In such a mixture, a solute is dissolved in another substance, known as a solvent. The solvent does the dissolving.- Types of solutions :...
in a smaller volume of eluent.
EBA may be considered to combine both the "Removal of Insolubles" and the "Isolation" steps of the 4-step downstream processing
Downstream processing
Downstream processing refers to the recovery and purification of biosynthetic products, particularly pharmaceuticals, from natural sources such as animal or plant tissue or fermentation broth, including the recycling of salvageable components and the proper treatment and disposal of waste. It is an...
heuristic. The major limitations associated with EBA technology is biomass interactions and aggregations onto adsorbent during processing.
Where classical column chromatography
Chromatography
Chromatography is the collective term for a set of laboratory techniques for the separation of mixtures....
uses a solid phase made by a packed bed, EBA uses particles in a fluidized
Fluidization
Fluidization is a process similar to liquefaction whereby a granular material is converted from a static solid-like state to a dynamic fluid-like state...
state. Expanded bed adsorption is, however, different from fluidised bed chormatography in essentially two ways: one, the EBA resin contains particles of varying size and density which results in a gradient of particle size when expanded; and two, when the bed is in its expanded state, local loops are formed. Particles such as whole cells or cell debris, which would clog a packed bed column, readily pass through a fluidized bed. EBA can therefore be used on crude culture broths or slurries of broken cells, thereby bypassing initial clearing steps such as centrifugation
Centrifugation
Centrifugation is a process that involves the use of the centrifugal force for the sedimentation of mixtures with a centrifuge, used in industry and in laboratory settings. More-dense components of the mixture migrate away from the axis of the centrifuge, while less-dense components of the mixture...
and filtration
Filtration
Filtration is commonly the mechanical or physical operation which is used for the separation of solids from fluids by interposing a medium through which only the fluid can pass...
, which is mandatory when packed beds are used. The feed flow rate is kept low enough that the solid packing remains stratified and does not fluidize completely. Hence EBA can be modeled as frontal adsorption
Adsorption
Adsorption is the adhesion of atoms, ions, biomolecules or molecules of gas, liquid, or dissolved solids to a surface. This process creates a film of the adsorbate on the surface of the adsorbent. It differs from absorption, in which a fluid permeates or is dissolved by a liquid or solid...
in a packed bed, rather than as a well mixed continuous-flow adsorber.
External links
- "Expanded-bed adsorption", at Modern Drug Discovery