299 IIUM Engineering Journal, Vol. 13 No. 1, 2012 Mohamed Safri et al. 97 PROCESS MODELLING AND DEBOTTLENECKING STUDY OF A VACCINE PRODUCTION NURUL HUDA MOHAMED SAFRI 1 , MAIZIRWAN MEL 1 , DOMINIC C.Y. FOO 2 , DENNY K.S. NG 2 AND IRENE M.L. CHEW 3 1 Bioprocess and Molecular Engineering Research Unit, Department of Biotechnology Engineering, Kulliyah of Engineering, International Islamic University Malaysia, P.O. Box 10, 50728 Kuala Lumpur, Malaysia. 2 Department of Chemical and Environmental Engineering, University of Nottingham Malaysia, Jalan Broga, 43500 Semenyih, Selangor, Malaysia 3 School of Engineering, Monash University Sunway Campus, Jalan Lagoon Selatan, 46150 Bandar Sunway, Selangor, Malaysia. maizirwan@iium.edu.my ABSTRACT: The main objective of this research work was to model and optimise the production of a locally-developed Infectious Coryza (IC) vaccine. The simulation work was performed using a commercially available batch process simulator SuperPro Designer v5.5. Six debottlenecking schemes were analysed using throughput analysis and cost to benefit ratio (CBR) when the annual production was set to increase by 100%. Based on the economic analysis, the selected debottlenecking scheme has an annual predicted revenue of USD 240 million, with a gross margin of 9.13% and a return on investment (ROI) of 46.12%. In addition, the payback period of the selected scheme is estimated to be within three years. ABSTRAK: Objektif utama dalam penyelidikan ini adalah untuk memodelkan dan mengoptimumkan hasil pembuatan vaksin tempatan Coryza berjangkit. Kerja simulasi ini dijalankan menggunakan alat simulasi Super Pro Designer v5.5. Sebanyak enam (6) skema khusus diujikaji menggunakan analisis pemprosesan dan kos kepada nisbah faedah (CBR) apabila pembuatan tahunan meningkat kepada 100%. Berdasarkan analisis ekonomi yang telah dilakukan, sesuatu skema khusus yang dipilih mempunyai keuntungan sebanyak USD 240 juta dengan margin kasar 9.13% dan pulangan atas pelaburan (ROI) sebanyak 46.12%. Selain itu juga, tempoh pembayaran balik bagi skema yang dipilih dianggarkan dalam tempoh tiga(3) tahun. KEYWORDS: process simulation; modelling; debottlenecking; optimisation 1. INTRODUCTION Malaysia is one of the countries with the highest chicken consumption per capita in the world at 32 kg. Some of the reasons for high chicken consumption are that chicken consumption is not against dietary prohibition or religious restrictions and chicken meat is a very low cost meat source in the country Malaysia Poultry and Products Annual 2006, 2006). Therefore, the poultry industry in Malaysia has grown from a backyard-type operation into a commercialised system in last 30 years. Due to the rapid grow of the poultry industry, the poultry diseases has poses a threat to the viability and productivity of poultry farming. IIUM Engineering Journal, Vol. 13 No. 1, 2012 Mohamed Safri et al. 98 Infectious Coryza (IC) disease is identified as one of the curses in the poultry industry worldwide [1]. IC is an acute respiratory disease of chickens which is caused by the bacterium known as Haemophilus paragallinarum (Hpg) [2]. Chickens of all ages are susceptible to this type of disease. Once the chickens are infected by this disease, the chickens get swollen eyes and nose, foul smelling discharges and sneezing. Besides, the chickens’ feed and water intake are reduced significantly which eventually leads to weight lost and lower production of egg [3]. In order to prevent this disease, the breeders in Malaysia normally administer vaccine which is imported from USA or Japan to the chickens [3]. However, the imported vaccines do not cure the infected chickens because the emergence of variant local strains of Hpg in the flocks [4]. In addition, the genetic makeup of local Hpg strains is different from the standard Hpg strains in the imported vaccines [5]. Hence, producing IC vaccine from the local Hpg strains is a better option to alleviate the IC disease problems in Malaysia [6]. With an effective local-made IC vaccine, the dependency of Malaysian breeders on the imported vaccines will be reduced. Process modelling and simulation in pharmaceutical industries functions as methodologies and tools that can be used to evaluate alternatives and speed up the development effort which may immensely impact on the bottom line [7]. With the given production capacity, modeling and simulation tool such as SuperPro Designer can be used to design and predict the feasibility of different production schemes [8]. For instance, Kumaresan et al. presented an approach to model and optimise Tongkat Ali extract process via SuperPro Designer [8]. Based on the study, a base case was first generated on the overall process and then debottlenecking strategies were proposed [8]. Besides that, detail economic analysis was also conducted. Based on the similar approach, modelling of IC vaccine production using the batch process simulation tool SuperPro Designer v5.5 is presented in this work. Due to the production capacity is limited by the current operating condition and equipment setup, debottlenecking study is performed to increase in annual production. Preliminary economic analysis is preformed to compare the debottlenecking schemes. Figure 1 shows the process flowsheet for a typical IC vaccine production designed with SuperPro Designer v5.5. The specifications for the major equipments used in the IC vaccine production are shown in Table 1. In order to produce IC vaccine through fermentation process, the bacteria Hpg is first prepared and transferred from a freezer (- 80°C) into a sterilized shake flask (P-1/SFR-101) contains media (nutrient and water). Note that the preparation of Hpg bacteria is excluded in this work. After 10 hours of pre- fermentation, the cultures were transferred to a 3 L seed fermentor (V-101), followed by a 30 L fermentor (V-103) and 300 L fermentor (V-104)., where the fermentation process is continued (see Fig. 1). It is worth mentioning that the media for the fermentation processes was pre-prepared in a media blending tank (V-102), and sterilized (in P-4 and P-6/ST-101) before it was transferred to the fermentors (V-103 and V-104) with the feed ratio of ten folds of the cultures feed. IIUM Engineering Journal, Vol. 13 No. 1, 2012 Mohamed Safri et al. 99 2. INFECTIOUS CORYZA (IC) VACCINE PRODUCTION P-1 / SFR-101 300 ml Inoculation P-2 / V-101 3 L Fermentation W ater 1 Hpg 1 W ater 2 Nutrient 1 Nutrient 2 P-5 / V-103 30 L Fermentation P-7 / V-104 300 L Fermentation P-3 / V-102 Media Blending Tank W ater Nutrient P-6 / ST-101 Heat Sterilization P-4 / ST-101 Heat Sterilization Media P-7 Media P-5 P-5 Media P-7 Media 3 L Hpg 30 L Hpg 300 ml Hpg BACTERIA FERMENTATION P-8 / DS-101 Centrifugation 300 L Hpg RECOVERY AND PURIFICATION PACKAGING Vent Out 1 Vent Out 2 Vent Out 3 Vent Out 4 Media P-9 / V-105 Kill Tank PBS Thimerosal Cell Paste P-11 / V-106 Reactor Tank Alum10% P-12 / HG-101 Homogenization Antigen+Adjuvan P-13 / V-107 Storage Vessel Mixture P-14 / FL-101 Filling Vial Vaccine Product P-10 / MF-101 Microfiltration Inactive Hpg Filtrate Concentrate Fig. 1 Process flowsheet of IC vaccine production (base case). Table 1: Major equipment specification for IC vaccine production. Quantity Procedure/ Equipment Specification Fermentation Section 1 P-1/SFR-101 Volume = 500 ml 1 P-2/V-101 Volume = 5 L 1 P-3/V-102 Volume = 330 L 1 P-4/ST-101 Rated throughput of 1080 L/h (Calculated based on design mode) 1 P-5/V-103 Volume = 50 L 1 P-6/ST-101 Rated throughput of 1080 L/h (Calculated based on design mode) 1 P-7/V-104 Volume = 500 L Recovery and Purification Section 1 P-8/DS-101 Based on Sigma factor 39627.55 m 2 (Calculated based on design mode) 1 P-9/V-105 Volume = 107.88 L (Calculated based on design mode) 1 P-10/MF-101 0.45 µ m membrane pore size 1 P-11/V-106 Volume = 77.96 L (Calculated based on design mode) 1 P-12/HG-101 Pumping efficiency of 70 % Packaging Section 1 P-13/V-107 Volume = 77.94 L (Calculated based on design mode) 1 P-14/FL-101 3000 entities/h (Calculated based on design mode) IIUM Engineering Journal, Vol. 13 No. 1, 2012 Mohamed Safri et al. 100 After the fermentation process, the cultures were harvested using a centrifuge (DS-101) and transferred to a kill tank (V-105) where the cultures were deactivated by adding thimerosal and phosphate buffer saline (PBS) [10]. Next, the cultures were further concentrated by passing it through a microfiltration membrane (MF-101) with pore size of 0.45 µ m and 99.7% removal efficiency, before it was being transferred to a reactor tank (V-106) [11]. Alum 10%, which acts as an adjuvant, was used an aid to the vaccine [12]. The adjuvant and antigen were homogenized in a homogenizer (HG-101) to ensure the same size particles. The mixture from the homogenizer was the final product and it was stored in a storage vessel (V-107) before it was sent for packing in filling machine (FL- 101).Tables 2 and 3 summarise the process scheduling (SUT: setup time; PT: process time; ST: start time) for each unit operation and the details of the raw materials (amount; price) in the IC vaccine production. 3. BOTTLENECK IDENTIFICATION STRATEGIES Based on the given information (Tables 1 – 3) and flowsheet in Fig.1, the process simulation of the based case is solved. The capital investment of the base case and the cost of production per unit are estimated as $18 million and $104.91, respectively. Based on the selling of $115 per unit, the annual revenue is computed as $123 million with 2.51 years of payback period. Table 2: Scheduling summary for operations and procedures in the base case model. Procedure/ Equipment Operation SUT (mins) PT ST P-1/SFR-101 Charge Nutrient 1 - 5 mins Beginning of batch Charge Water 1 - 3 mins After Nutrient 1 charge Agitation - 5 mins After Water 1 charge Charge Hpg 1 - 3 mins After Agitation Fermentation - 10 hours After Hpg 1 charge Transfer out 300 mL Hpg to P-2 - 3 mins After Fermentation P-2/V-101 Charge Nutrient 2 - 5 mins After 12 hours of batch operation Charge Water 2 - 3 mins After Nutrient 2 charge Agitation - 8.4 mins After Water 2 charge Transfer in 300 mL Hpg from P-1 - Master-Slave with P-1 Transfer Out 300 mL Hpg Starts with Transfer 300 ml in P-1 (to P-2) Fermentation - 6 hours After Transfer in 300 mL Hpg from P-1 Transfer out 3 L Hpg to P-5 - 3 mins After Fermentation CIP - 15 mins After Transfer out to P-3 IIUM Engineering Journal, Vol. 13 No. 1, 2012 Mohamed Safri et al. 101 Table 3 (Continued): Scheduling summary for operations and procedures in the base case model. Procedure/ Equipment Operation SUT (mins) PT ST P-3/V-102 Charge Nutrient 20 Calculated based on 600 L/h volumetric flowrate After 14.3 hours of batch operation Charge Water 20 Calculated based on 600 L/h volumetric flowrate After Charge Nutrient Agitation - 10 mins After Charge Water Transfer Out Media P-5 to P-4 20 Calculated based on 600 L/h volumetric flowrate After Agitation Store - 4.66 hours After Transfer Out Media P-5 Transfer Out Media P-7 to P-6 20 Calculated based on 600 L/h volumetric flowrate After Store CIP - 15 mins After Transfer Out Media P-7 P-4/ST-101 Sterilize - 15 mins Starts with Transfer Out Media P-5 in P-3 (to P-4) P-5/V-103 Transfer In Media P-5 from P-4 20 Calculated based on 600 L/h volumetric flowrate After Sterilize in P-4 Transfer In 3 L Hpg from P-2 - Master Slave with P-2 Transfer Out 3 L Hpg Starts with Transfer Out 3 L Hpg in P-2 (to P-5) Fermentation - 5 hours After Transfer In 3 L Hpg Transfer Out 30 L Hpg to P-7 20 Calculated based on 600 L/h volumetric flowrate After Fermentation CIP - 15 mins After Transfer Out 30 L Hpg P-6/ST-101 Sterilize - 15 mins Starts with Transfer Out Media P-7 in P-3 (to P-6) P-7/V-104 Transfer In Media P-7 from P-6 20 Calculated based on 600 L/h volumetric flowrate After Sterilize in P-6 Transfer In 30 L Hpg from P-5 - Master Slave with P-5 Transfer Out 30 L Hpg Starts with Transfer Out 30 L Hpg in P-5 (to P-7) Fermentation - 4.5 hours After Transfer In 30 L Hpg Transfer Out Broth to P- 8 - Master Slave with P-8 Centrifuge After Fermentation CIP - 15 mins After Transfer Out Broth IIUM Engineering Journal, Vol. 13 No. 1, 2012 Mohamed Safri et al. 102 Table 4 (Continued): Scheduling summary for operations and procedures in the base case model. Procedure/ Equipment Operation SUT (mins) PT ST P-8/DS-101 Centrifuge - 60 mins Starts with Transfer Out Broth in P-7 (to P-8) CIP - 15 mins After Centrifuge P-9/V-105 Transfer In Cell Paste from P-8 - Master Slave with P-8 Centrifuge Starts with Centrifuge in P-8 Charge PBS - Calculated based on 600 L/h flow rate After Transfer In Cell Paste Agitation - 10 mins After Charge PBS Charge Thimerosal - Calculated based on 600 L/h flow rate After Agitation Agitation - 15 mins After Charge Thimerosal Transfer out Inactive Hpg to P-10 20 Calculated based on 600 L/h flow rate After Agitation CIP - 15 mins After Transfer Out Inactive Hpg to P-10 P-10/MF-101 Filtration - 120 mins Starts with Transfer Out Inactive Hpg in P-9 (to P- 10) CIP - 15 mins After Filtration P-11/V-106 Transfer in Concentrate from P-10 20 Master Slave with P-10 Filtration Starts with Filtration in P- 10 Charge Alum10% - Calculated based on 600 L/h flow rate After Transfer In Concentrate Agitation - 15 mins After Charge Alum10% Transfer Out Antigen+Adjuvant to P- 12 20 Calculated based on 600 L/h flow rate After Agitation CIP - 15 mins After Transfer Out Antigen+Adjuvant P-12/HG-101 Homogenize - 30 mins Starts with Transfer Out Product Antigen+Adjuvant in P-11 (to P-12) CIP - 15 mins After Homogenize P-13/V-107 Transfer in Mixture from P-12 20 Calculated based on 600 L/h flow rate Starts with Homogenize in P-12 Transfer Out Product to P-13 20 Calculated based on 600 L/h flow rate After Transfer In Mixture CIP - 15 mins After Transfer Out Product P-14/FL-101 Filling (Fill level: 50 mL/bottle) 20 Master Slave with P-13 Transfer Out Product Starts with Transfer Out Product in P-13 (to P-14) (SUT: setup time; PT: process time; ST: start time) IIUM Engineering Journal, Vol. 13 No. 1, 2012 Mohamed Safri et al. 103 Table 5: Raw materials used in a single batch Raw Materials Symbol Approximation Amount (kg/batch) Price (USD/kg) Aluminum hydroxide Alum10% Aluminum oxide 7.867 2.00 Chicken Serum Chicken Serum Protein 8.481 207.15 Glucose Glucose Glucose 50.886 25.14 Haemophillusparagallinarum Hpg Biomass 0.032 0.00 Disodium hydrogen phosphate Na2HPO4 Sodium hydrogen phosphate 25.443 85.14 NADH NADH Protein 1.696 75,000.00 Phosphate buffer saline PBS Sodium chloride 38.701 14.29 Peptone Peptone Protein 50.886 128.00 Sodium chloride Sodium Chloride Sodium chloride 32.228 8.57 Thimerosal Thimerosal Ethyl benzene 0.013 10.17 Water Water Water 7,318.739 0.02 As the increase of the demand of IC, the management decided to increase the production capacity by 100%. Therefore, the current process is facing difficulties to meet the requirement. In order to overcome the problem, the process bottleneck is first identified. Based on the set target, throughput analysis is first performed to identify the process bottlenecks, i.e. scheduling or size bottlenecks. Generally, bottleneck could be caused by the limitation of equipment or resources such as utilities, labor and raw materials supply. Based on the identified bottleneck, different debottlenecking schemes are proposed, and economic analysis is carried out for selection of the scheme with highest CBR. Note that CBR is defined as the ratio of extra benefit to the extra cost as shown in Equation (1) [7]. ��� � ���� �� ���� ���� � ��� � ���� (1) In order to increase the annual process throughput to 100%, three strategies are considered, i.e. increase of batch size (the amount of product produced per batch of operation), increase number of batches or increase of both batch size and number of batches. According to Petrides et al., the scheduling bottleneck can be identified by tracking the total time consumed by each equipment within its cycle time [7]. The equipment with the longest cycle time is identified as the scheduling bottleneck, and this bottleneck will determine the maximum number of batches [12]. On the other hand, the size bottleneck of a process can be determined by calculating the capacity utilisation, uptime and combined utilisation of the various processing steps such as fermentation, centrifugation and filtration [10]. Capacity utilisation of equipment is referred to the fraction of equipment capacity that is used during an operation. Meanwhile, uptime is defined as ratio of equipment’s occupancy time over the plant cycle time. Combined utilisation is the product of the capacity used and the uptime. This parameter clearly shows the time and capacity of particular equipment that is being used over the process. IIUM Engineering Journal, Vol. 13 No. 1, 2012 Mohamed Safri et al. 104 Fig. 2 The operation Gantt Chart (base case). Figure 2 shows the operation Gantt Chart of the base case. As shown in Fig. 2, it is noted that shake flask (SFR-101) is identified as the scheduling bottleneck due to its longest occupancy (10 hours) as compare to the other equipment. The next step is to identify the debottlenecking schemes as shown in the following section. 4. DEBOTTLENECKING SCHEMES TO INCREASE PRODUCTION After process bottleneck is identified, six debottlenecking schemes are proposed to increase the annual production of IC vaccine to 100%. As presented previously, shake flask (SFR-101) is identified as the scheduling bottleneck; hence, in order to increase the annual production, debottlenecking strategies should target to reduce the fermentation time in the shake flask. In order to reduce the fermentation time of shake flask (SFR-101), another set of 300 mL shake flask (SFR-102) that staggered the operation is considered. By staggering the shake flask the overall cycle time of shake flask is reduced by half. In this scheme (scheme 1), the annual throughput increased 39.48% as compared to the base case (1067 batches per year), while the CBR value is calculated at 0.74. This scheme provides a good debottlenecking alternative as the cost of shake flask is very low compared with the overall capital investment. Since the annual throughput is only increased by 39.48%, the process needs to be further debottlenecked in order to achieve the targeted 100% increment. Throughput analysis is next carried out for Scheme 2, media bleeding tank (P3/V-102) is identified as the next scheduling bottleneck. Hence, an additional media blending tank that operates in staggered mode is added. The annual throughput of this scheme is increased to 1136 batches, i.e. 48.5% increment from the base case. Meanwhile, the CBR is determined as 0.77 which is slightly higher than Scheme 1. IIUM Engineering Journal, Vol. 13 No. 1, 2012 Mohamed Safri et al. 105 The throughput analysis is repeated on Scheme 2 to generate Scheme 3. In Scheme 3 an additional 5 L fermentor (V-109) is added to operate in staggered mode with the current fermentor (V-101), after Schemes 1 and 2 are considered. The annual throughput of this scheme is increased to 1141 batches, i.e. 49.2% increment from the base case. Note that, the annual number of batches does not increase much as compare with Scheme 2. Meanwhile, the CBR of this scheme is same as Scheme 2. Thus, this scheme is not as attractive as compared to Scheme 2. In Scheme 4, throughput analysis reveals that the 500 L fermentor (V-104) emerges as the new process bottleneck due to its longest occupancy time due to its master slave relationship with centrifugation process and its long fermentation time. Hence, an extra set of 500 L fermentor is added. Based on the simulated result, the annual throughput for this scheme is increased to 1223 batches or 59.9% increment, and the CBR of this scheme is determined as 0.79. The 50 L fementor (V-103) is next identified as the new process bottleneck for Scheme 4. In order to eliminate this process bottleneck, an additional 50 L fermentor is installed (Scheme 5). The simulated result shows that the number of batches tremendously increased to 1492 and 95.03% increment from the base case, and the CBR is calculated as 0.86. It is worth noting that the CBR for Scheme 5 is the highest CBR among all five debottleneck schemes. However, its annual production is yet to reach to 100%. BACTERIA FERMENTATION P-1 / SFR-101 300 ml Inoculation P-2 / V-101 3 L Fermentation Water 1 Hpg 1 Water 2 Nutrient 1 Nutrient 2 P-5 / V-103 30 L Fermentation P-7 / V-104 300 L Fermentation P-3 / V-102 Media Blending Tank Water Nutrient P-6 / ST-101 Heat Sterilization P-4 / ST-101 Heat Sterilization Media P-7 P-5 Media P-7 Media 30 L Hpg (5) P-8 / DS-101 Centrifugation 300 L Hpg RECOVERY AND PURIFICATION PACKAGING Vent Out 1 Vent Out 2 Vent Out 3 Vent Out 4 Media P-9 / V-105 Kill Tank PBS Thimerosal Cell Paste P-11 / V-106 Reactor Tank Alum10% P-12 / HG-101 Homogenization Antigen+Adjuvan P-13 / V-107 Storage Vessel Mixture P-14 / FL-101 Filling Vial Vaccine Product P-10 / MF-101 Microfiltration Inactive Hpg Filtrate Concentrate P-1A / SFR-102 New Shake Flask Nutrient 1A Water 1A Hpg 1A 300 ml Hpg (1) P-2A / V-108 New 5 L Fermentor Nutrient 2A Water 2A300 ml Hpg (1A) 3 L Hpg (2) P-3A / V-109 New Media Blending Tank Nutrient A Water A P-5A / V-110 New 50 L Fermentor P-7A / V-111 New 500 L Fermentor P-6A / ST-102 New Heat Sterilization P-4A / ST-103 New Heat Sterilization S-101 P-7A Media S-103 P-5A Media S-105 S-106 30 L Hpg (5A) Fig. 3 Debottlenecking Scheme 6. Scheme 6 is introduced where an additional heat steriliser is installed (see Fig. 3). This equipment is operated in staggered mode with the existing heat steriliser. The annual throughput increases to 1530 batches from 765 batches in the base case. Note that 100% increment of annual production is achieved. Even though the CBR value for Scheme 6 is computed as 0.85, which is slightly lower than that in Scheme 5, this scheme fulfils the IIUM Engineering Journal, Vol. 13 No. 1, 2012 Mohamed Safri et al. 106 objective of the debottlenecking study. The debottlenecking process is a continuous process as there is always a limitation of equipment for the overall production. After completing Scheme 6, it is found that shake flask (P1/SFR-101) is once again becomes the new process bottleneck as it is now the longest process that limits the process cycle time. Although debottlenecking is a continuous process, it stops when the scheme that achieves the company's target is achieved. For instance, in this case study, debottlenecking is stopped at Scheme 6 as it met the 100% production rise as compared to the base case. Table 4 shows the summary of all debottlenecking schemes as well as the base case. Table 6: Economic comparison of the base case study and debottlenecking strategy Scenario % production increase (Annual batches) Annual Throughput (vials) Cost of investment ($) Annual Operating Cost ($) Annual Revenue ($) Unit Production Cost ($/vial) CBR Base case 0 % (765) 1,074,395.354 18,202,250 112,710,857 123,555,466 104.9063 - Scheme 1 39.5% (1067) 1,498,535.742 22,184,430 156,633,492 172,331,610 104.5244 0.74 Scheme 2 48.5% (1136) 1,595,441.990 23,429,706 166,728,627 183,475,829 104.5031 0.77 Scheme 3 49.2 % (1141) 1,602,464.182 23,761,950 167,502,583 184,283,381 104.5281 0.77 Scheme 4 59.9% (1223) 1,717,628.128 26,573,045 179,740,333 197,527,235 104.6445 0.79 Scheme 5 95.0% (1492) 2,095,422.050 30,785,330 218,980,377 240,973,546 104.5042 0.86 Scheme 6 100% (1530) 2,148,790.708 32,938,306 224,803,395 247,110,931 104.6186 0.85 5. CONCLUSION In this work, process simulation tool is used to model and simulate the IC vaccine production. 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