Scale up of industrial microbial processes
- 1. SCALE UP OF INDUSTRIAL MICROBIAL PROCESSES (IM 601) SUBMITTED TO JANE CLARYN BENJAMIN MA’AM SUBMITTED BY (GROUP IV) SHUBHAM RANJAN 18BTFT034 KULDEEP GEHLOT 18BTFT035 ADITI PATIDAR 18BTFT036 SMRITI SINGH 18BTFT038 MANISHA SALONI 18BTFT041 PRASHANT SAINI 18BTFT042 SMITA KUMARI 18BTFT043 KAKUMANU SANJYOPTHA 18BTFT045 TANYA VATS 18BTFT048
- 2. TABLE OF CONTETS 1. INTRODUCTION 2. WHAT IS SCALE UP AND WHY DOES IT MATTER 3. STAGES OF SCALE UP FERMENTATION 4.KEY CONSIDERATION FOR FERMENTATION SCALE UP 5. ROAD MAP FOR SUCCESSFUL SCALE-UP 6.KEY CHALLENGES AND IMPROVEMENT IN MICROBIAL SCALE UP PROCESSES 7. FERMENTATION PROCESS SCALE-UP INCLUDE 8.WHAT ARE THE OPPORTUNITIES IN SCALE UP OF INDUSTRIAL MICROBIAL PROCESS CONCLUSION
- 3. 1. INTRODUCTION The importance of biotechnological production is increasing all over the world. It’s importance increases as it is connected with health and food industry directly or indirectly. So overall it is a billion dollar industry. The way these impressive economic biotech figures can be achieved is either to develop completely new bioproducts and to successfully launch them on the market or to expand existing production capacities against the competition of market forces. The concerted approach of metabolic engineering, synthetic biology and systems biology is promising to develop optimized processes in lab-scale, they still need to be transferred into large-scale without reduction of sensitive performance parameters.
- 4. Notably, large-scale performance losses are not unusual. The explanation of scale-up mechanisms for preventing unwanted performance losses is not only an academic goal but also an economic necessity. Microbial fermentation processes play a critical role in many important industrial applications. In various applications, fermentation processes are utilized to produce final product, convert substrates, catalyze reactions or simply make mass biological materials. This project covers a lot of information related to scale up industrial microbial processes like principles for process scale up, key considerations for fermentation scale up, road map, challenges, improvements and opportunities.
- 5. 2. WHAT IS SCALE UP AND WHY DOES IT MATTER The successful startup and operation of commercial size unit is design and operating procedure are in the path based upon experimentation and demonstration it is smaller scale. Scale-up is increasing something in size, amount, or production. Microbial processes involve cultivation of microbes in bioreactors (also referred to as fermentors) to produce a product, as well as the subsequent recovery and purification of the product and disposal of associated wastes. Scale-up of microbial processes is undertaken typically for a commercial purpose, specifically to provide product benefits to customers and to generate a financial return for investors.
- 6. It serves to minimize the risk of a large capital investment in the full-scale manufacturing plant by further validating the process, the supply chain (from raw materials to commercial product application), and market demand If the degree of novelty is low, then the demonstration plant may be skipped. For the remainder of the article, we will use ‘pilot’ in reference to both pilot and demo scales. The financial investment to scale up a microbial process to manufacturing scale is usually greater than the cost to develop the production microbe and lab-scale process The financial risk is high, so deterioration in process performance during scale-up will be costly and disruptive, potentially even leading to project failure. So, in scaling up microbial processes, it is clearly impactful to get it right and to get it right the first time.
- 7. 3. STAGES OF SCALE UP FERMENTATION Scale-up of large industrial processes is preferably done in two stages if there is a high degree of novelty in the process and/or the commercial product. The first stage is a pilot plant (pilot scale) with 100–10,000 L fermenters and matched downstream equipment. Its purpose is to translate the lab-scale process into a realistic scaled-down version of the manufacturing process. In most cases, the process is not fully integrated; i.e. each individual unit operation is operated batch-wise. The second stage of scale-up is a demonstration plant (demo scale) with 10 000–100 000 L fermenters and matched downstream. It serves to minimize the risk of a large capital investment in the full-scale manufacturing plant by further validating the process, the supply chain and market demand.
- 8. 3.1 Guiding principles for process scale up The authors have contributed to the commercialization of a variety of industrial microbial processes, including first-of-a-kind projects, from early stage R&D to scale-up to manufacturing support . There are three guiding principles that are critical to the successful scale-up of industrial microbial processes. 1.Begin with the end in mind The challenge is having a realistic and accurate view of what the end looks like especially when it’s a first-of-a-kind process. One cannot simply enlarge lab-scale equipment and duplicate lab scale conditions at large-scale.
- 9. Without an understanding of large-scale equipment and how scale dependent parameters change, a project is likely to get into big trouble. Instead, beginning with the end in mind, a skilled project team that really understands large scale processes prepares a detailed conceptual design of the envisioned manufacturing process and plant before the first lab experiments are done. Use the conceptual design to provide early guidance to the experimental R&D program on process viability and key scale and economic parameters. Then regularly update the design as your experimental program produces new learnings.
- 10. 2. Be diligent in the details Unfortunately, we’ve seen all kinds of oversights and shortcuts during process scale-up, with consequences ranging from disruptive to catastrophic. On the other hand, with close attention given to critical details, microbial processes can be scaled up with minimal unpleasant surprises. Ultimately, this will reward stakeholders with a safe, reliable manufacturing plant that meets or exceeds its financial objectives.
- 11. 3. Prepare for the unexpected Common examples include utility interruptions, microbial contamination, variable raw material quality, fouling of process equipment, equipment failure and unexpected poor process performance at scale. Prioritize based on risk magnitude and prepare a detailed risk mitigation plan. For high magnitude risks relating to process upsets, design lab/pilot studies to assess the impact on process performance and develop a detailed process upset response plan to inform the plant operations team of the proper mode of action if an upset does occur.
- 12. 4.KEY CONSIDERATION FOR FERMENTATION SCALE UP Poor fermentation performance at large-scale is almost always considered a priority scale-up risk. This is because fermentation is usually the costliest process step, both in terms of variable costs (raw materials and utilities) and capital investment. Fermentation is also a complex unit operation. There are many parameters that impact performance, and most of these are subject to change during scale-up. A. Preparation of inoculum B. Tank/Fermenter Aspect Ratio C. Sterilization and cleaning of the tank D. Proper supply of oxygen and agitation E. Control of foaming
- 13. A. Preparation of inoculum – is the first and most important parameter because the inoculum size, density and the phase in which the inoculum is will determine the yield and productivity.
- 14. B. Tank/Fermenter Aspect Ratio – the capacity of the fermenter in the Industries used are large . So the size and capacity of the fermenter should be kept in mind while applying the other methods and processes.
- 15. C. Sterilization and cleaning of the tank – the tank and other equipments should be cleaned and sterilized to ensure the protection of media inside the fermenter from any contamination D. Proper supply of oxygen and agitation – The demand for oxygen in SUF is always more and it is necessary to keep the level of oxygen more than the critical level and the supplied oxygen and air should be thoroughly mixed inside the fermenter E. Control of foaming – during the process foams generated might spill out and may contaminate the Product inside the fermenter so antifoaming agent should be used. F. Other factors such as pH, removal of heat and pressure
- 16. 5. ROAD MAP FOR SUCCESSFUL SCALE-UP The scale-up journey starts by imagining the desired outcome a robust full-scale manufacturing plant that meets its commercial objectives (schedule, cost, and quality) begin with the ending mind. All of this is remembered at the beginning of a project in the form of a detailed, written charter that is updated as the program progresses from R&D proof-of-concept through process development and eventually deployment. But in the case of fermentation, large-scale conditions can usually be adequately simulated in stirred lab fermenters with the implementation of Some custom control hardware and software.
- 17. A fully integrated process, including recycle streams, can be operated for an extended period with fully representative industrial equipment and materials. Alternative equipment designs and suppliers can be evaluated. The future large-scale plant operating team can be trained; from the team’s pilot experience, they will know the process works and will log valuable experience in addressing process upsets. Pilot plant data and operating know-how are used to improve the large-scale plant design. Large quantities of product can be produced for customer evaluation in the end-use applications, which builds customer relationships, confidence and demand for the commercial plant output.
- 18. As a Results, it can be tempting to skip this step or to pilot only selected unit operations for a short time. Experience has proven this to be unwise; the downside far outweighs the modest upside. Design of the large-scale manufacturing plant should be based on data generated in the pilot plant. It is also important to factor in the outlook for future technology improvements. But avoid the temptation to design the large-scale plant for a process that has not been validated at pilot scale. Whether you intend to build, own and operate your own plant or license your technology, it is crucial to provide technincal support during all phases of the project, including engineering design, construction, commissioning and start-up.
- 20. 6.KEY CHALLENGES AND IMPROVEMENT IN MICROBIAL SCALE UP PROCESSES Regardless of how well you prepare, there will be issues that arise during scale-up. Common examples include utility interruptions, microbial contamination, variable raw material quality, fouling of process equipment, equipment failure and unexpected poor process performance at scale. Improvements mean that the focus now is not so much on capacity as on purification. The big challenges now lie in downstream processing. “There are lots of ideas out there,”. In both downstream processing and in other stages of manufacture, scale up can present a challenge but one that can be met by employing the right technologies.
- 21. Scale up generally involves taking a lab-scale procedure and replicating it as closely as possible to get larger amounts of product as specified either by a client or the regulatory authorities typically in a fermenter.
- 22. 6.1 SOME OF THE ISSUSES OFFERMENTATION SCALE UP PROCESSES CHROMATOGRAPHY The use of chromatography in purification can lead to some challenges in scale up, which may be solved by new approaches. Mixed-mode Hypercell media can exploit different mechanisms of interaction with a target protein. FILTRATION Filtration is necessary at many stages of a manufacturing process, so an understanding of filterability the filtration behavior of a solution is crucial. Filterability testing can be used to determine the capacity of a filter system with the aim of determining the filter area that would be needed to process a batch without blocking the filter upon scale up.
- 23. HYDROCYCLONE Many industries have been looking at a new cell separation technology based upon the Hydrocyclone, which can be used as a perfusion system in large-scale animal cell culture. The hydrocyclone works like an inverted fixed-wall centrifuge without a rotating shaft.
- 24. 7. FERMENTATION PROCESS SCALE-UP INCLUDE 7.1 Seed train sale-up strategy The purpose of a seed train is to propagate cells to a desired mass for inoculation into the production bioreactor. The traditional seed train includes thawing a vial and inoculating into shaker flasks for a certain number of stages with increasing flask size, and may include stainless steel reactors. The guiding principle for a seed train scale up is to minimize the number of stages for entire seed train, maintain robust growth and finally prevent any negative impact of extended generations to productivity and quality.
- 25. Typical studies of seed train scale-up may include:- 1.Vial thaw conditions: this may include temperature and time between thaw and inoculation 2.Inoculation ratio: for microbes,0.1to 5% is commonly used. However, overall process robustness needs to be demonstrated with the inoculation ratios. 3.Seed train media: Usually, seed train media are optimized to support growth, rather than product expression, in a batch mode. 4.Genetic stability and production stability during the seed train process: This requires studies with an extended duration, beyond that estimated to be required for generation at the production scale.
- 27. 7.2 Production fermentation scale-up The essence of scale-up of a fermentation process is to demonstrate fermentation production at large scale resulting in the same productivity and quality as that developed at small scale. One of the outcomes of a process scale-up is to finalize a detailed large scale process description with settings of all operational parameters and their ranges at scale. The scaled-up fermentation has to be demonstrated with a certain number of runs for consistency and statistical significance. To achieve these goals, one have to take the following steps Identify linear scale-up parameters Temperature Ph Pressure Dissolved oxygen(DO) Air flow rate
- 28. 8.WHAT ARE THE OPPORTUNITIES IN SCALE UP OF INDUSTRIAL MICROBIAL PROCESS Opportunities in scale up of Industrial microbial process Many researches were done and the analysis showed potential target genes associated with increased ATP demand for metabolic engineering. Such approach was previously used by the same research team for re‐designing an E. coli strain with increased glucose uptake capacity to cope with nutrient‐limiting conditions typically found in industrial fed‐batch bioprocess . These new findings point out that predicting microbial physiological responses to environmental fluctuations remains a challenge but also that there are vast potential applications.
- 29. This then is a good opportunity for presenting a modernized version of the scale‐down approach, i.e. an outlook on the different items that have to be considered for a better understanding of bioprocess scale‐up and the behaviour of cell populations under industrial process‐related conditions . The bioprocess industry more than ever needs new, efficient and sustainable routes to manufacture bio‐products. Bioprocesses use the power and versatility of nature via microorganisms that make the bio‐products from renewable feedstocks. These microorganisms today can be extensively re‐programmed into efficient cell factories.
- 31. CONCLUSION Scaling up industrial microbial processes for commercial production is a high stakes endeavor ,requiring time and investment often exceeding that for laboratory microbe and process development. The bio‐economy is in transit from innovation to commercialization. The bioprocess industry is expected to increasingly deliver bio‐products to the market, in large amounts, at high quality and at competitive cost levels. This requires flawless start‐up of new large‐scale bioprocesses and continuous improvement of running processes.
- 32. Process scale up ,in a broad sense, is a critical activity that enables a fermentation process achieved in research and development to operate at a commercially viable scale for manufacturing .A successful scale up involves many aspects of successful preparation and planning beyond pure process scale up technology.