- Email: [email protected]
Development of bioprocessing for Vero cell derived JE vaccine
Abstracts / Journal of Biotechnology 136S (2008) S201–S211
6000 (6-12%) and the results were compared with those from acid precipitation method using phosphoric acid. Although the ratio of HBsAg/total protein was higher using acid precipitation but the amount of detected antigen doubled when 10% PEG 6000 was applied. Keywords: Clarification; Pichia pastoris; Hepatitis B surface antigen; Polyethylene glycol; Acid precipitation doi:10.1016/j.jbiotec.2008.07.441 III5-P-017 Aluminum phosphate shows more adjuvanticity than aluminum hydroxide in recombinant hepatitis B vaccine formulation Mohammad Reza Fazeli 1,∗ , Rassoul Dinarvand 1 , Nasrin Samadi 1 , Arash Mahboubi 1 , Hooshmand Ilka 2 , Mohammad Sharifzadeh 3 , Saeed Azadi 4 , Ali Moghanlou 4 , Mohammad Mirzaei Salehabady 4 , Mahboubeh Valadkhani 5 1
Department of Drug and Food Control, Faculty of Pharmacy, Tehran University of Medical Sciences, Iran 2 Zist Daru Danesh Ltd., Iran 3 Department of Toxicology and Pharmacology, Faculty of Pharmacy, Tehran University of Medical Sciences, Iran 4 Biotechnology Unit, Daru Pakhsh Manufacturing Company, Iran 5 Department of Biological Products, Ministry of Health, Iran E-mail address: [email protected] (M.R. Fazeli).
Although lots of researches have been carried out to find alternative adjuvants to aluminum salts in vaccine formulations, they are still extensively used due to their good track record of safety, low cost and proper adjuvanticity with a variety of antigens. Adsorption of antigens onto aluminum compounds depends heavily on electrostatic forces between adjuvant and antigen. Commercial recombinant protein hepatitis B vaccines containing aluminum hydroxide as adjuvant is facing low induction of immunity in some sections of the vaccinated population. To follow the current global efforts in finding more potent hepatitis B vaccine formulation, adjuvanticity of aluminum phosphate has been compared to aluminum hydroxide. The adjuvant properties of aluminum hydroxide and aluminum phosphate in a vaccine formulation containing a locally manufactured hepatitis B (HBs) surface antigen was evaluated in Balb/c mice. The formulations were administered inter peritoneum (i.p.) and the titers of antibody induced after 28 days were determined using ELISA technique. The geometric mean of antibody titer (GMT), seroconversion and seroprotection rates, ED50 and relative potency of different formulations were determined. All the adjuvanicity markers obtained in aluminum phosphate formulation were significantly higher than aluminum hydroxide. The geometric mean of antibody titer of aluminum phosphate was approximately three folds more than aluminum hydroxide. Aluminum phosphate showed more adjuvanticity than aluminum hydroxide in hepatitis B vaccine. Therefore the use of aluminum phosphate as adjuvant in this vaccine may lead to higher immunity with more durable effect in vaccinated groups. doi:10.1016/j.jbiotec.2008.07.442
S209
III5-P-020 Development of bioprocessing for Vero cell derived JE vaccine Soo-Jin Moon, Ik-Hwan Kim ∗ School of Life Sciences and Biotechnology, Korea University, SeongbukGu, Seoul 136-701, Republic of Korea E-mail address: [email protected] (I.-H. Kim). The vaccine of Japanese Encephalitis Virus (JEV) is produced in the brain of mouse or fertilized eggs currently (Sugawara et al., 2002). However the conventional processes have many problems, such as the high cost of production, the controversy over bioethics for sacrificing animals and the impurities derived from mouse brain may cause neurological diseases (Sugawara et al., 2002; Bharati and Vrati, 2006). These problems could be resolved by the production system for JE vaccine derived from Vero cells with serum free media (Sugawara et al., 2002; Qiesney et al., 2003). In this work, the optimum condition of JE vaccine production is established and the assay titrating JEV for rapid detection would be developed. Furthermore, to increase the production of JE vaccine, the development of scaleup system with bioreactor and the effect of addition of antioxidant would be examined. The Vero cell growth kinetics and infectivity of JEV strain against the Vero cells were determined by measuring cell density and JEV infectious titers. The immunogenicity of Vero cell-derived JEV was determined by ELISA method with an Eantigen. The specific protein of JEV was detected by using western blotting. In order to determine the high production conditions of JE vaccine, various parameters for Vero cell culture with microcarriers and virus infection in a bioreactor would be investigated. References Bharati, K., Vrati, S., 2006. Japanese encephalitis: development of new candidate vaccines. Expert Rev. Anti. Infect. Ther. 4, 313–324. Qiesney, S., Marc, A., Gerdil, C., Gimenez, C., Marvel, J., Richard, Y., Meignier, B., 2003. Kinetics and metabolic specificities of Vero cells in bioreactor cultures with serum-free medium. Cytotechnology 42, 1–11. Sugawara, K., Nishiyama, K., Ishikawa, Y., Abe, M., Sonoda, K., Komatsu, K., Horikawa, Y., Takeda, K., Honda, T., Kuzuhara, S., Kino, Y., Mizokami, H., Mizuno, K., Oka, T., Honda, K., 2002. Development of Vero cell-derived inactivated Japanese encephalitis vaccine. Biologicals 30, 303–314.
doi:10.1016/j.jbiotec.2008.07.443 III5-P-021 Prediction of the HLA class I binding consensus peptides for influenza A (H5N1) Linh Thuoc Tran ∗ , Hai Van Van, Thi Thanh Thuy Le, Thi Ngoc Phuong Cao University of Natural Sciences, Vietnam National University, Ho Chi Minh City, Viet Nam E-mail address: [email protected] (L.T. Tran). Influenza A (H5N1) viruses could cause severe worldwide epidemic, with high attack rates, large numbers of deaths. Effective vaccines against these viruses in humans are urgently needed (Horimoto and Kawaoka, 2006). One major problem is their limitation to the specific H5N1 groups. Therefore, we built a system predicting the HLA class I binding consensus peptides on viral envelope proteins towards multivalent epitope-based vaccines against challenge infection with different groups of H5N1 (Tamar and Ruth,
∗ Corresponding author. Tel.: +82 2 3290 3447.
6000 (6-12%) and the results were compared with those from acid precipitation method using phosphoric acid. Although the ratio of HBsAg/total protein was higher using acid precipitation but the amount of detected antigen doubled when 10% PEG 6000 was applied. Keywords: Clarification; Pichia pastoris; Hepatitis B surface antigen; Polyethylene glycol; Acid precipitation doi:10.1016/j.jbiotec.2008.07.441 III5-P-017 Aluminum phosphate shows more adjuvanticity than aluminum hydroxide in recombinant hepatitis B vaccine formulation Mohammad Reza Fazeli 1,∗ , Rassoul Dinarvand 1 , Nasrin Samadi 1 , Arash Mahboubi 1 , Hooshmand Ilka 2 , Mohammad Sharifzadeh 3 , Saeed Azadi 4 , Ali Moghanlou 4 , Mohammad Mirzaei Salehabady 4 , Mahboubeh Valadkhani 5 1
Department of Drug and Food Control, Faculty of Pharmacy, Tehran University of Medical Sciences, Iran 2 Zist Daru Danesh Ltd., Iran 3 Department of Toxicology and Pharmacology, Faculty of Pharmacy, Tehran University of Medical Sciences, Iran 4 Biotechnology Unit, Daru Pakhsh Manufacturing Company, Iran 5 Department of Biological Products, Ministry of Health, Iran E-mail address: [email protected] (M.R. Fazeli).
Although lots of researches have been carried out to find alternative adjuvants to aluminum salts in vaccine formulations, they are still extensively used due to their good track record of safety, low cost and proper adjuvanticity with a variety of antigens. Adsorption of antigens onto aluminum compounds depends heavily on electrostatic forces between adjuvant and antigen. Commercial recombinant protein hepatitis B vaccines containing aluminum hydroxide as adjuvant is facing low induction of immunity in some sections of the vaccinated population. To follow the current global efforts in finding more potent hepatitis B vaccine formulation, adjuvanticity of aluminum phosphate has been compared to aluminum hydroxide. The adjuvant properties of aluminum hydroxide and aluminum phosphate in a vaccine formulation containing a locally manufactured hepatitis B (HBs) surface antigen was evaluated in Balb/c mice. The formulations were administered inter peritoneum (i.p.) and the titers of antibody induced after 28 days were determined using ELISA technique. The geometric mean of antibody titer (GMT), seroconversion and seroprotection rates, ED50 and relative potency of different formulations were determined. All the adjuvanicity markers obtained in aluminum phosphate formulation were significantly higher than aluminum hydroxide. The geometric mean of antibody titer of aluminum phosphate was approximately three folds more than aluminum hydroxide. Aluminum phosphate showed more adjuvanticity than aluminum hydroxide in hepatitis B vaccine. Therefore the use of aluminum phosphate as adjuvant in this vaccine may lead to higher immunity with more durable effect in vaccinated groups. doi:10.1016/j.jbiotec.2008.07.442
S209
III5-P-020 Development of bioprocessing for Vero cell derived JE vaccine Soo-Jin Moon, Ik-Hwan Kim ∗ School of Life Sciences and Biotechnology, Korea University, SeongbukGu, Seoul 136-701, Republic of Korea E-mail address: [email protected] (I.-H. Kim). The vaccine of Japanese Encephalitis Virus (JEV) is produced in the brain of mouse or fertilized eggs currently (Sugawara et al., 2002). However the conventional processes have many problems, such as the high cost of production, the controversy over bioethics for sacrificing animals and the impurities derived from mouse brain may cause neurological diseases (Sugawara et al., 2002; Bharati and Vrati, 2006). These problems could be resolved by the production system for JE vaccine derived from Vero cells with serum free media (Sugawara et al., 2002; Qiesney et al., 2003). In this work, the optimum condition of JE vaccine production is established and the assay titrating JEV for rapid detection would be developed. Furthermore, to increase the production of JE vaccine, the development of scaleup system with bioreactor and the effect of addition of antioxidant would be examined. The Vero cell growth kinetics and infectivity of JEV strain against the Vero cells were determined by measuring cell density and JEV infectious titers. The immunogenicity of Vero cell-derived JEV was determined by ELISA method with an Eantigen. The specific protein of JEV was detected by using western blotting. In order to determine the high production conditions of JE vaccine, various parameters for Vero cell culture with microcarriers and virus infection in a bioreactor would be investigated. References Bharati, K., Vrati, S., 2006. Japanese encephalitis: development of new candidate vaccines. Expert Rev. Anti. Infect. Ther. 4, 313–324. Qiesney, S., Marc, A., Gerdil, C., Gimenez, C., Marvel, J., Richard, Y., Meignier, B., 2003. Kinetics and metabolic specificities of Vero cells in bioreactor cultures with serum-free medium. Cytotechnology 42, 1–11. Sugawara, K., Nishiyama, K., Ishikawa, Y., Abe, M., Sonoda, K., Komatsu, K., Horikawa, Y., Takeda, K., Honda, T., Kuzuhara, S., Kino, Y., Mizokami, H., Mizuno, K., Oka, T., Honda, K., 2002. Development of Vero cell-derived inactivated Japanese encephalitis vaccine. Biologicals 30, 303–314.
doi:10.1016/j.jbiotec.2008.07.443 III5-P-021 Prediction of the HLA class I binding consensus peptides for influenza A (H5N1) Linh Thuoc Tran ∗ , Hai Van Van, Thi Thanh Thuy Le, Thi Ngoc Phuong Cao University of Natural Sciences, Vietnam National University, Ho Chi Minh City, Viet Nam E-mail address: [email protected] (L.T. Tran). Influenza A (H5N1) viruses could cause severe worldwide epidemic, with high attack rates, large numbers of deaths. Effective vaccines against these viruses in humans are urgently needed (Horimoto and Kawaoka, 2006). One major problem is their limitation to the specific H5N1 groups. Therefore, we built a system predicting the HLA class I binding consensus peptides on viral envelope proteins towards multivalent epitope-based vaccines against challenge infection with different groups of H5N1 (Tamar and Ruth,
∗ Corresponding author. Tel.: +82 2 3290 3447.