Downstream processing
Encyclopedia
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 essential step in the manufacture of pharmaceuticals such as antibiotics, hormones (e.g. insulin and human growth hormone), antibodies (e.g. infliximab and abciximab) and vaccines; antibodies and enzymes used in diagnostics; industrial enzymes; and natural fragrance and flavor compounds. Downstream processing is usually considered a specialized field in biochemical engineering
, itself a specialization within chemical engineering
, though many of the key technologies were developed by chemists and biologists for laboratory-scale separation of biological products.
Downstream processing and analytical bioseparation both refer to the separation or purification of biological products, but at different scales of operation and for different purposes. Downstream processing implies manufacture of a purified product fit for a specific use, generally in marketable quantities, while analytical bioseparation refers to purification for the sole purpose of measuring a component or components of a mixture, and may deal with sample sizes as small as a single cell.
Removal of insolubles is the first step and involves the capture of the product as a solute in a particulate-free liquid, for example the separation of cells
, cell debris or other particulate matter from fermentation broth containing an antibiotic. Typical operations to achieve this are filtration
, centrifugation
, sedimentation
, precipitationflocculation
, electro-precipitation, and gravity settling. Additional operations such as grinding, homogenization, or leaching, required to recover products from solid sources such as plant and animal tissues, are usually included in this group.
Product isolation is the removal of those components whose properties vary markedly from that of the desired product. For most products, water is the chief impurity and isolation steps are designed to remove most of it, reducing the volume of material to be handled and concentrating the product. Solvent
extraction, adsorption
, ultrafiltration, and precipitation
are some of the unit operations involved.
Product purification is done to separate those contaminants that resemble the product very closely in physical and chemical properties. Consequently steps in this stage are expensive to carry out and require sensitive and sophisticated equipment. This stage contributes a significant fraction of the entire downstream processing expenditure. Examples of operations include affinity, size exclusion, reversed phase chromatography
, crystallization
and fractional precipitation.
Product polishing describes the final processing steps which end with packaging of the product in a form that is stable, easily transportable and convenient. Crystallization
, desiccation
, lyophilization and spray drying are typical unit operations. Depending on the product and its intended use, polishing may also include operationsto sterilize the product and remove or deactivate trace contaminants which might compromise product safety. Such operations might include the removal of viruses
or depyrogenation
.
A few product recovery methods may be considered to combine two or more stages. For example, expanded bed adsorption
(Vennapusa et al. 2008) accomplishes removal of insolubles and product isolation in a single step. Affinity chromatography
often isolates and purifies in a single step.
Fermentation (food)
Fermentation in food processing typically is the conversion of carbohydrates to alcohols and carbon dioxide or organic acids using yeasts, bacteria, or a combination thereof, under anaerobic conditions. Fermentation in simple terms is the chemical conversion of sugars into ethanol...
broth, including the recycling of salvageable components and the proper treatment and disposal of waste. It is an essential step in the manufacture of pharmaceuticals such as antibiotics, hormones (e.g. insulin and human growth hormone), antibodies (e.g. infliximab and abciximab) and vaccines; antibodies and enzymes used in diagnostics; industrial enzymes; and natural fragrance and flavor compounds. Downstream processing is usually considered a specialized field in biochemical engineering
Biochemical engineering
Biochemical engineering is a branch of chemical engineering or biological engineering that mainly deals with the design and construction of unit processes that involve biological organisms or molecules, such as bioreactors...
, itself a specialization within chemical engineering
Chemical engineering
Chemical engineering is the branch of engineering that deals with physical science , and life sciences with mathematics and economics, to the process of converting raw materials or chemicals into more useful or valuable forms...
, though many of the key technologies were developed by chemists and biologists for laboratory-scale separation of biological products.
Downstream processing and analytical bioseparation both refer to the separation or purification of biological products, but at different scales of operation and for different purposes. Downstream processing implies manufacture of a purified product fit for a specific use, generally in marketable quantities, while analytical bioseparation refers to purification for the sole purpose of measuring a component or components of a mixture, and may deal with sample sizes as small as a single cell.
Stages in downstream processing
A widely recognized heuristic for categorizing downstream processing operations divides them into four groups which are applied in order to bring a product from its natural state as a component of a tissue, cell or fermentation broth through progressive improvements in purity and concentration.Removal of insolubles is the first step and involves the capture of the product as a solute in a particulate-free liquid, for example the separation of cells
Cell (biology)
The cell is the basic structural and functional unit of all known living organisms. It is the smallest unit of life that is classified as a living thing, and is often called the building block of life. The Alberts text discusses how the "cellular building blocks" move to shape developing embryos....
, cell debris or other particulate matter from fermentation broth containing an antibiotic. Typical operations to achieve this are 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...
, 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...
, sedimentation
Sedimentation
Sedimentation is the tendency for particles in suspension to settle out of the fluid in which they are entrained, and come to rest against a barrier. This is due to their motion through the fluid in response to the forces acting on them: these forces can be due to gravity, centrifugal acceleration...
, precipitationflocculation
Flocculation
Flocculation, in the field of chemistry, is a process wherein colloids come out of suspension in the form of floc or flakes by the addition of a clarifying agent. The action differs from precipitation in that, prior to flocculation, colloids are merely suspended in a liquid and not actually...
, electro-precipitation, and gravity settling. Additional operations such as grinding, homogenization, or leaching, required to recover products from solid sources such as plant and animal tissues, are usually included in this group.
Product isolation is the removal of those components whose properties vary markedly from that of the desired product. For most products, water is the chief impurity and isolation steps are designed to remove most of it, reducing the volume of material to be handled and concentrating the product. Solvent
Solvent
A solvent is a liquid, solid, or gas that dissolves another solid, liquid, or gaseous solute, resulting in a solution that is soluble in a certain volume of solvent at a specified temperature...
extraction, 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...
, ultrafiltration, and precipitation
Precipitation (chemistry)
Precipitation is the formation of a solid in a solution or inside anothersolid during a chemical reaction or by diffusion in a solid. When the reaction occurs in a liquid, the solid formed is called the precipitate, or when compacted by a centrifuge, a pellet. The liquid remaining above the solid...
are some of the unit operations involved.
Product purification is done to separate those contaminants that resemble the product very closely in physical and chemical properties. Consequently steps in this stage are expensive to carry out and require sensitive and sophisticated equipment. This stage contributes a significant fraction of the entire downstream processing expenditure. Examples of operations include affinity, size exclusion, reversed phase chromatography
Chromatography
Chromatography is the collective term for a set of laboratory techniques for the separation of mixtures....
, crystallization
Crystallization
Crystallization is the process of formation of solid crystals precipitating from a solution, melt or more rarely deposited directly from a gas. Crystallization is also a chemical solid–liquid separation technique, in which mass transfer of a solute from the liquid solution to a pure solid...
and fractional precipitation.
Product polishing describes the final processing steps which end with packaging of the product in a form that is stable, easily transportable and convenient. Crystallization
Crystallization
Crystallization is the process of formation of solid crystals precipitating from a solution, melt or more rarely deposited directly from a gas. Crystallization is also a chemical solid–liquid separation technique, in which mass transfer of a solute from the liquid solution to a pure solid...
, desiccation
Desiccation
Desiccation is the state of extreme dryness, or the process of extreme drying. A desiccant is a hygroscopic substance that induces or sustains such a state in its local vicinity in a moderately sealed container.-Science:...
, lyophilization and spray drying are typical unit operations. Depending on the product and its intended use, polishing may also include operationsto sterilize the product and remove or deactivate trace contaminants which might compromise product safety. Such operations might include the removal of viruses
Viral processing
The main idea behind viral processing is to stop the viruses in a given sample from infecting the desired product. The two most widely used methods of viral processing are viral removal and viral inactivation. The former is a method in which all viruses are simply removed from the sample completely...
or depyrogenation
Depyrogenation
Depyrogenation refers to the removal of pyrogens from solution, most commonly from injectable pharmaceuticals.A pyrogen is defined as any substance that can cause a fever. Bacterial pyrogens include endotoxins and exotoxins, although many pyrogens are endogenous to the host...
.
A few product recovery methods may be considered to combine two or more stages. For example, expanded bed adsorption
Expanded bed adsorption
Expanded bed adsorption is a preparative chromatographic technique which makes processing of viscous and particulate liquids possible.-Principle:...
(Vennapusa et al. 2008) accomplishes removal of insolubles and product isolation in a single step. 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:...
often isolates and purifies in a single step.
See also
- Separation processSeparation processIn chemistry and chemical engineering, a separation process, or simply a separation, is any mass transfer process used to convert a mixture of substances into two or more distinct product mixtures, at least one of which is enriched in one or more of the mixture's constituents. In some cases, a...
- Unit operationUnit operationIn chemical engineering and related fields, a unit operation is a basic step in a process.Unit operation involves bringing a physical change such as separation, crystallization, evaporation, filtration etc.. For example in milk processing, homogenization, pasteurization, chilling, and packaging are...
- Fermentation (biochemistry)Fermentation (biochemistry)Fermentation is the process of extracting energy from the oxidation of organic compounds, such as carbohydrates, using an endogenous electron acceptor, which is usually an organic compound. In contrast, respiration is where electrons are donated to an exogenous electron acceptor, such as oxygen,...
- Validation (drug manufacture)Validation (drug manufacture)Validation in the pharmaceutical, medical device, food, blood establishments, tissue establishments, and clinical trials industries is defined as the documented act of demonstrating that a procedure, process, and activity will consistently lead to the expected results. It often includes the...