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This article has been written by Dr. Yogita Lugani pursuing a Diploma in Intellectual Property: Prosecution, licensing and litigation from LawSikho.

Introduction

Scaling up bioprocesses is a critical attempt in biotechnology and microbiology, aiming to transfer technology from a research laboratory to commercial scale after detecting its commercial viability. This process involves the optimisation of parameters at the flask level and comparing these results on a large scale for industrial production by exploring the challenges and developing strategies for scaling-up of bioprocesses. The ultimate goal of scaling-up is to achieve improved production, maintain the quality of the product and determine the economic feasibility of the process. Microorganisms are diverse in nature and prove potential for the synthesis of various industrial products like alcohols, amino acids, antibiotics, beverages, enzymes, pharmaceutical drugs, vaccines, vitamins and organic acids. The microbial isolates that show industrial potential are identified using various morphological, physiological, biochemical, chemotaxonomic and molecular techniques. After identification, the isolates are submitted to a microbial depository/ gene bank for long-term preservation using glycerol stocks, lyophilization and cryopreservation methods. The novel isolates are generally deposited under a safe deposit or patent deposit to get commercial benefit from these strains. The technologies using natural isolates for commercial gains need prior approval from the National Biodiversity Authority with a benefit-sharing agreement. There are many regulatory approvals and quality controls before marketing the product.

Microbial products of industrial importance

Microorganisms prove tremendous applications for the synthesis of various industrial products, including primary and secondary metabolites synthesised at log and stationary phases, respectively. Microorganisms can survive in diverse environments, including low temperatures, specifically -20 °C (psychrophiles), high temperatures of 80 °C (thermophiles), and extremely high temperatures of up to 110 °C (hyper-thermophiles). Similar to temperature, microbes can also thrive well under acidic conditions with a pH of 2 (acidophile) and alkaline conditions with a pH of 11 (alkalophile). Extremophiles, i.e., microbial cells that can grow and produce primary and secondary metabolites at high temperatures, acidic pH and high osmotic concentrations, are of industrial importance. Microbial cells that require oxygen for growth are called aerobes, whereas microbes that can grow well under oxygen deficient conditions are called anaerobes. Therefore, companies are spending millions of dollars in their research and development (R&D) for the isolation of potential microbial candidates that can be exploited for the synthesis of industrial products. The microbially synthesised products, which are of industrial importance, include amino acids, antibiotics, biofuels, food additives, enzymes, organic acids, vaccines, pharmaceutical drugs, beverages, single cell proteins, vitamins, biofertilizers, etc. Microbes are a good source for producing natural drugs possessing antiviral, antimicrobial, immunosuppressive and cytotoxic activities. 

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Tailoring of media and process parameters at flask level

Industrial products that include microbiological and fermentation processes utilise microbial strains, for which there is a need for tailoring/ or customisation of various media and process parameters to enhance the production yield, ensure product consistency and reduce the production cost of the desired product. Media parameters involve the carbon source with its concentration, the nitrogen source with its composition and combination of organic and inorganic nitrogen sources, trace elements that may act as inducers or inhibitors for various enzyme-based reactions, osmolarity to prevent cell stress and induce specific metabolic pathways. Process parameters include the effects of temperature, pH, aeration and agitation rate, dissolved oxygen, pressure and gas composition. Utilisation of agro-industrial wastes such as wheat straw, rice straw, paddy straw, bagasse, and corn cob, which are available in huge amounts in developing countries like India, further helps to reduce the cost of production along with promoting the circular economy principle.

Statistical methods, algorithms, and modelling approaches, including Design of Experiments, Response Surface Methodology, Multivariate Data Analysis, Artificial Intelligence, Machine Learning, and Fluid Dynamics, help to understand and optimise the media and process parameters in industrial applications.

Scaling-up of bioprocesses for commercialisation

Scaling-up refers to shifting from laboratory scale to commercial scale, and it includes both upsteam and downstream processes with the final aim of launching a product in the market. Scaling-up of bioprocesses is a complex process that requires interdisciplinary collaboration between biologists, engineers, chemists, regulatory experts, and business professionals to ensure the successful commercialization of biotechnological products. It takes into account bioreactor design, cell culture and production conditions at the fermentor level, harvesting and recovery of the product, quality control and regulatory compliance associated with the product to be marketed, validation studies, risk assessment and contingency plans, market and economic considerations, and continuous improvement with optimisation.

The media and process parameters, which have been optimised at the flask level, may not work well at the bioreactor level due to scale effects, mass transfer limitations, cell viability, shear stress and the time required for inoculum adaptation. Hence, understanding the metabolic pathway, cellular kinetics, and critical process parameters (CPPs) helps in the detailed characterization of the process. Scaling factors such as aeration rate, agitation size, oxygen transfer coefficient, heat dissipation, and vessel size need to be determined based on empirical data and engineering principles. The feed rate of the media depends on the type of fermentation process, i.e.

  • In batch fermentation, in which all the media components are added only once, followed by product recovery.
  • In fed-batch fermentation, media is added in small intervals to achieve better product yield.
  • In continuous fermentation, there is the continuous addition of media containing nutrients and inducers from the inlet and removal of the product from the outlet to avoid the accumulation of inhibitors and toxic substances.

To make production cost-effective, most of the industrial processes are conducted on a fed-batch and continuous scale.

Downstream processing is done for product recovery by separating biomass, purifying and concentrating. Purification of the product is carried out by using different filtration, chromatography, centrifugation, precipitation, extraction, dialysis and ultrafiltration techniques.

Validation studies are required during the scale-up of bioprocesses to ensure product quality, process consistency and reliability at the industrial scale. Identification of potential risks like equipment failure, contamination of media and the development of contingency plans during commercial operations are other parameters that need to be considered during the commercial synthesis of products.

Factors considering during technology transfer

There are various factors that need to be considered while transferring knowledge, skills, know-how or intellectual property from one organisation to another. The patent status of a technology is very important, as it is quite easy to transfer technologies for which patents have already been granted in different jurisdictions, including India. Whereas, the technologies for which patent applications that have been either abandoned or rejected by the Contoller General of Patents, Designs and Trademarks are difficult to transfer and commercialise. Technology Readiness Level (TRL) is very crucial during the valuation of a technology, i.e., a technology with a low TRL (initial level technology) is expected to be transferred at a lower amount because there are a lot of efforts and inputs required by the recipient entity; however, a technology with a high TRL (advanced level technology) is valued at a higher amount as this technology is almost ready and minimum inputs are required from the recipient entity before commercialising the product. The economic viability of a technology is estimated by its initial investment, operational costs, potential revenues, and return/ profit from the investment. Scaling up bioprocesses from the laboratory to commercial production is a critical step in the development of new biopharmaceuticals and other biologics. This process involves a number of challenges, including:

  • Maintaining product quality and consistency
  • Ensuring process efficiency and cost-effectiveness
  • Meeting regulatory requirements

To successfully scale up a bioprocess, a number of practical approaches can be employed. These approaches include:

  • Process characterisation: This involves understanding the key process parameters that affect product quality and yield. This information can be used to design a scale-up strategy that minimises the risk of process failure.
  • Pilot-scale studies: These studies are conducted at a smaller scale than commercial production, but they are large enough to provide meaningful data on process performance. Pilot-scale studies can be used to optimise process conditions and identify potential problems that may arise during scale-up.
  • Process validation: This involves demonstrating that the scaled-up process consistently produces a product that meets all of the required specifications. Process validation is typically conducted through a series of validation runs, which are designed to test the process under a variety of operating conditions.
  • Technology transfer: This involves transferring the scaled-up process from the development laboratory to the commercial manufacturing facility. Technology transfer is a complex process that requires careful planning and execution.

By following these practical approaches, bioprocess companies can successfully scale up their processes and bring new products to market.

Additional considerations for scaling up bioprocesses

In addition to the practical approaches described above, there are a number of other factors that should be considered when scaling up a bioprocess. These factors include:

  • Regulatory compliance: Bioprocess companies must ensure that their scaled-up processes comply with all applicable regulatory requirements. This includes obtaining the necessary permits and licenses, and conducting the required environmental impact assessments.
  • Cost of scale-up: The cost of scaling up a bioprocess can be significant. This cost includes the cost of building or expanding a manufacturing facility, the cost of purchasing new equipment, and the cost of training personnel.
  • Time to market: Bioprocess companies must balance the need to scale up their processes quickly with the need to ensure product quality and regulatory compliance. A rushed scale-up can lead to problems that can delay the launch of a new product.

By carefully considering all of these factors, bioprocess companies can successfully scale up their processes and bring new products to market in a timely and cost-effective manner.

The technical feasibility of the technology is another parameter that is very crucial, and it considers all the resources and existing infrastructure of the recipient entity and checks the viability of the technology. Training, skill development and support are necessary to build capability and capacity in the recipient organisation. Understanding market demand and opportunity by considering market size, competitive products available in the market, consumer preference and regulatory requirements helps to understand market demand and potential commercialization prospects of the technology. Risk assessment includes all the risks associated with market acceptance, implementation challenges and legal issues.

Considering all these factors, a successful transfer of technology requires a maximum gain in profits and long term good relationship between the provider and recipient entities.

Regulatory approvals

While transferring the products or technologies originating from nature and/or natural resources of the Indian Territory from Research and Development (R&D) to the industrial level, various regulatory approvals need to be obtained. If the research possess any biological material which is isolated directly from the natural environment or is derived from the natural isolate, National Biodiversity Approval (NBA) is required under Biodiversity Act, 2002 by filing Form-II. Similarly, National Biofuel Policy of India promotes the use of biofuels, also called as green fuel, in India to fulfil the enhancing fuel demand of rising population and prevent environmental pollution. Any entity with biofuel production facility needs to obtain permission from National Biofuel Authority (NBFA) to ensure compliance with environmental, safety and quality standards. Permission for environmental clearance is obtained from Ministry of Environment, Forest and Climate Change (MoEFCC).

The fermentation products that are projected for human consumption, such as nutraceuticals, health drinks, organic acids, amino acids, etc., need permission from the Food Safety and Standards Authority of India (FSSAI). The pharmaceutical products, vaccines and drugs require clearance from the Drug Controller General of India (DCGI) after conducting various preclinical and clinical trials. These products can be marketed only after getting approval by DCGI to meet safety and efficacy standards.       

Conclusion

It has been concluded that microorganisms are industrially important biocatalysts, and they can be isolated from diverse habitats. These isolates can be exploited for the synthesis of various industrial products. Various media and process parameters are tailored at the flask level to achieve maximum product yield. Shifting from the lab/pilot scale to a commercial scale involves product synthesis at the bioreactor level, where some parameters like aeration rate, agitation rate, and oxygen transfer coefficient need to be optimised. Following product synthesis, there is product recovery by downstream processing with the final aim of enhancing the purity of the product. The parameters that need to be considered while transferring technology from one entity to another are intellectual property status, technology readiness level, technical feasibility in the recipient entity and economic viability of the technology. Different regulatory approvals need to be obtained from the authority before the launch of biotechnological products into the market.

References

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