Removing residual DNA from Vero-cell culture-derived human rabies vaccine by using nuclease |
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| Institution: | 1. Harbin University of Commerce, Harbin 150028, China;2. School of Life Science, Northeast Agricultural University, Harbin 150030, China;3. Lubin Environmental Equipment Protection Co., Ltd, Shanghai 200125, China;4. University of Chinese Academy of Sciences, Beijing 100049, China;1. National Bioproducts Institute, Pinetown, South Africa;2. Graduate Institute of Biomedical Materials and Tissue Engineering, Taipei Medical University, 250 Wuxing St., Taipei City 110, Taiwan;1. iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2780-901 Oeiras, Portugal;2. Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, Portugal;3. EMD Millipore Corporation, 80 Ashby Road, Bedford, MA 01730, USA;1. Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901 Oeiras, Portugal;2. Instituto de Tecnologia Química e Biológica, Universidade NOVA de Lisboa, 2780-157 Oeiras, Portugal;3. LAQV-REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal;4. Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade NOVA de Lisboa, 2829-516 Caparica, Portugal;5. Sartorius Stedim Biotech GmbH, Spindler-Strasse11, 37079 Gottingen, Germany;1. Oswaldo Cruz Foundation (FIOCRUZ), Bio-Manguinhos, Avenida Brasil 4365, 21045-900 Rio de Janeiro/RJ, Brazil;2. Federal University of Rio de Janeiro (UFRJ), Institute of Chemistry, Biochemistry Program, Av. Athos da Silveira Ramos 149 – bl. A, 21941-909 Rio de Janeiro/RJ, Brazil;3. Federal University of Rio de Janeiro (UFRJ), Institute of Microbiology, Virology Department, Av. Carlos Chagas 373 – bl. I, 21941-902 Rio de Janeiro/RJ, Brazil;4. Federal University of Rio de Janeiro (UFRJ), COPPE, Cell Culture Engineering Laboratory, Cx. Postal 68502, 21941-972 Rio de Janeiro/RJ, Brazil |
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| Abstract: | The clearance of host cell DNA is a critical indicator for Vero-cell culture-derived rabies vaccine. In this study, we evaluated the clearance of DNA in Vero-cell culture-derived rabies vaccine by purification process utilizing ultrafiltration, nuclease digestion, and gel filtration chromatography. The results showed that the bioprocess of using nuclease decreased residual DNA. Dot-blot hybridization analysis showed that the residual host cell DNA was <100 pg/ml in the final product. The residual nuclease in rabies vaccine was less than 0.1 ng/ml protein. The residual nuclease could not paly the biologically active role of digestion of DNA. Experiments of stability showed that the freeze-drying rabies virus vaccine was stable and titers were >5.0 IU/ml. Immunogenicity test and protection experiments indicated mice were greatly induced generation of neutralizing antibodies and invoked protective effects immunized with intraperitoneal injections of the rabies vaccine. These results demonstrated that the residual DNA was removed from virus particles and nuclease was removed by gel filtration chromatography. The date indicated that technology was an efficient method to produce rabies vaccine for human use by using nuclease. |
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| Keywords: | Vero-cell culture-derived rabies vaccine Residual host cell DNA Nuclease digestion Gel filtration chromatography |
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