Question Description: What are the inputs to and outputs from Downstream Processing operations for a conventional biotechnological process?
Course Hero Answer & Explanation:
- Downstream processing is process of recovering and purifying the products obtained in the procedure. The process includes series of events.
Inputs of the process
- The success of the downstream process depends on the inputs used in the protocol and the parameters maintained in the procedure.
- In a industrial level synthesis of a product depends on regulation and monitoring of many factors.
- For example: Factors to be monitored for microbiologically synthesized product
Screening and selection of the organism
- Depends on the product synthesized in the process the microbial cells are selected. In general bacteria, yeast, fungi and mammalian cell lines are the mostly used in the process.
- Bacteria yields a higher productivity, resistance to heat damage, osmotic pressure and availability
- Yeast also demonstrate higher productivity, high cell concentrations, easy maintenance of culture. Nowadays , genetically engineered strains are much useful in synthesis of tailor made products
- Selection of the organism or cell line plays a vital role in as they have a huge impact on the scale of production and feeding regimes such as batch, fed-batch, continuous and perfusion culture and quality of the product
Formulating the media required for the production
- The formulation of growth medium is ideally should contain the essential nutrients that supports the growth of the microbes.
- There are two types of media are useful in fermentation process such as inoculum media and production media that enhances the growth of the organism initially and serve as source of carbon and nitrogen for the further growth of the organism throughout the process.
Optimization of media
- The optimized media is selected depends on the quality and quantity of the product synthesized.
- Raw materials such as cellulose, corn syrup, sugar, beet and corn molasses serve as carbon sources for the fermentation procedure
Process control requirements such as pH, temperature
- Monitoring and maintenance of pH and temperature throughout the process play a paramount role as the smaller deviation of these factors drastically affect the quality and yield of the product.
- Use of acid and base and thermostat function are suggested to regulate these important factors.
Outputs of the process
There are five stages in the downstream processing which can be listed as,
Separation of biomass-product
- The separation is attempted by disruption of cell and harvesting
- In this step the cellular biomass is harvested by removing the particulate matter from the fermentation broth which is carried out by centrifugation or by filtration techniques.
Discharge of Intracellular products
- The release of the intracellular products such as vitamins and enzymes in the active form are achieved by the lysis of the cell.
- The cell disruption can be achieved by physical methods such as ultrasonication, heat shock, applying high pressure, using the glass beads for grinding, chemical methods such as use of organic solvents, detergents and alkalies, and enzymatic methods by using enzymes such as lysozyme, protease etc.
- The appropriate selection of the method depends on the product synthesized in the procedure.
Concentration of the products
- In this step the product is obtained in a purified form by eliminating the contaminants and concentrating the products in homogenous form.
Application of chromatography by purification
- This step plays a prominent role in the purification of the product by utilizing various types of chromatography and filtration techniques.
- The fermented products such as proteins, pharmaceuticals and research components are effectively purified by this technique that separates the compound from the mixture of compounds. Gel filtration, Ion exchange, affinity, hydrophobic interaction and immobilized metal-ion affinity are the various chromatography techniques employed in this step based on the nature of the product to be purified.
Formulation of the product
- In this step the activity and stability purified product is maintained by following proper storage and addition of stabilizing agents. For example, proteins and vitamins are highly susceptible to loss of stability, hence addition of ideal stabilizing agents are highly important.
- Some of the stabilizers used in the protein formulation re sucrose, sodium chloride, polyethylene glycol and glycerol. The products are marketed in preferable forms such as dry powders, solutions etc.
- The downstream processing is a crucial procedure in product development, which plays a major role in industrial scale in biotechnological and pharmaceutical category.
- Synthesis of biological or chemicals are imperative for numerous applications counting diagnostics, and for research and development industry.
- The strategy to be followed during the downstream processing plays a main role in the successful yield of the products.
- The use of purified products in the various industrial applications makes the maintenance of quality during the downstream processing is of chief importance.
- The different stages of handling that happen after the completion of the fermentation include partition of the biomass , refinement by filtration and chromatography, and formulation of the products by addition of stabilizing agents.
For example : vitamins, antibiotics and proteins used in pharmaceutical industry