title: Removing residual DNA from Vero-cell culture-derived human rabies vaccine by using nuclease.
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.
authors.affiliation: Harbin University of Commerce, Harbin 150028, China; School of Life Science, Northeast Agricultural University, Harbin 150030, China.
authors.forename: Si-Ming
authors.initials: SM
authors.lastname: Li
authors.affiliation: School of Life Science, Northeast Agricultural University, Harbin 150030, China; Lubin Environmental Equipment Protection Co., Ltd, Shanghai 200125, China.
authors.forename: Fu-Liang
authors.initials: FL
authors.lastname: Bai
authors.affiliation: School of Life Science, Northeast Agricultural University, Harbin 150030, China.
authors.forename: Wen-Juan
authors.initials: WJ
authors.lastname: Xu
authors.affiliation: School of Life Science, Northeast Agricultural University, Harbin 150030, China.
authors.forename: Yong-Bi
authors.initials: YB
authors.lastname: Yang
authors.affiliation: School of Life Science, Northeast Agricultural University, Harbin 150030, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: An
authors.affiliation: University of Chinese Academy of Sciences, Beijing 100049, China.
authors.forename: Tian-He
authors.initials: TH
authors.lastname: Li
authors.affiliation: School of Life Science, Northeast Agricultural University, Harbin 150030, China.
authors.forename: Yin-Hang
authors.initials: YH
authors.lastname: Yu
authors.affiliation: School of Life Science, Northeast Agricultural University, Harbin 150030, China.
authors.forename: De-Shan
authors.initials: DS
authors.lastname: Li
authors.affiliation: School of Life Science, Northeast Agricultural University, Harbin 150030, China. Electronic address: wenfeiwang_bio@126.com.
authors.forename: Wen-Fei
authors.initials: WF
authors.lastname: Wang
mesh_terms: D000818:Animals; D057134:Antibodies, Neutralizing / Q000096:biosynthesis; D000914:Antibodies, Viral / Q000096:biosynthesis; D002522:Chlorocebus aethiops; D002850:Chromatography, Gel; D004247:DNA / Q000302:isolation & purification*; D004340:Drug Contamination / Q000517:prevention & control; D004355:Drug Stability; D004706:Endodeoxyribonucleases / Q000302:isolation & purification; D004722:Endoribonucleases / Q000302:isolation & purification; D005612:Freeze Drying; D006801:Humans; D051379:Mice; D011818:Rabies / Q000276:immunology / Q000517:prevention & control; D011819:Rabies Vaccines / Q000276:immunology / Q000302:isolation & purification*; D011820:Rabies virus / Q000276:immunology; D014709:Vero Cells
chemical_list: D057134:Antibodies, Neutralizing; D000914:Antibodies, Viral; D011819:Rabies Vaccines; D004247:DNA; D004706:Endodeoxyribonucleases; D004722:Endoribonucleases; C021835:Serratia marcescens nuclease
keywords: Gel filtration chromatography; Nuclease digestion; Residual host cell DNA; Vero-cell culture-derived rabies vaccine