title: Development a scalable production process for truncated human papillomavirus type-6 L1 protein using WAVE Bioreactor and hollow fiber membrane.
abstract: Here, we describe a process for expression, purification, and characterization of truncated human papillomavirus type-6 (HPV-6) L1 virus-like particles (VLPs). The scalable cultivation process in a WAVE Bioreactor at the 10-L scale was optimized to express HPV-6 L1 VLPs using the baculovirus insect expression system. A hollow fiber membrane system was used for the integrated operation, including concentration, diafiltration, extraction, and clarification. The HPV-6 L1 protein was further purified by anion-exchange chromatography and hydrophobic chromatography. The HPV-6 L1 protein could self-assemble into VLPs with a diameter of approximately 50-60 nm after removal of the reductant dithiothreitol (DTT). The final purified HPV-6 L1 VLPs product was characterized to estimate yield and purity, and exceeds the requirements for pharmaceutical-grade VLP vaccine. Immunization of mice demonstrated that the vaccine could elicit high titer neutralizing antibodies in vivo. This study confirms the feasibility of producing pharmaceutical-grade HPV type-6 L1 VLPs on an industrial scale for clinical trials.
authors.affiliation: Nation Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun, 130012, China.
authors.forename: Bo
authors.initials: B
authors.lastname: Sun
authors.affiliation: Nation Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun, 130012, China.
authors.forename: Dandan
authors.initials: D
authors.lastname: Zhao
authors.affiliation: Nation Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun, 130012, China.
authors.forename: Xizhen
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Nation Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun, 130012, China.
authors.forename: Tiejun
authors.initials: T
authors.lastname: Gu
authors.affiliation: Nation Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun, 130012, China.
authors.forename: XiangHui
authors.initials: X
authors.lastname: Yu
authors.affiliation: Nation Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun, 130012, China.
authors.forename: Shiyang
authors.initials: S
authors.lastname: Sun
authors.affiliation: Nation Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun, 130012, China.
authors.forename: Xinghong
authors.initials: X
authors.lastname: Zhao
authors.affiliation: Jilin Medical University, Jilin City, Jilin Province, 132013, China.
authors.forename: Liu
authors.initials: L
authors.lastname: Wei
authors.affiliation: BCHT Biotechnology Company, Changchun, 130012, China.
authors.forename: Dawei
authors.initials: D
authors.lastname: Liu
authors.affiliation: BCHT Biotechnology Company, Changchun, 130012, China.
authors.forename: Hui
authors.initials: H
authors.lastname: Yan
authors.affiliation: Nation Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun, 130012, China.
authors.forename: Xiangyu
authors.initials: X
authors.lastname: Meng
authors.affiliation: Nation Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun, 130012, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Kong
authors.affiliation: Nation Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun, 130012, China.
authors.forename: Fei
authors.initials: F
authors.lastname: Xu
authors.affiliation: BCHT Biotechnology Company, Changchun, 130012, China.
authors.forename: Ping
authors.initials: P
authors.lastname: Yang
authors.affiliation: Nation Engineering Laboratory for AIDS Vaccine, Jilin University, Changchun, 130012, China. jiangcl@jlu.edu.cn.
authors.forename: Chunlai
authors.initials: C
authors.lastname: Jiang
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics / Q000378:metabolism; D019149:Bioreactors; D036022:Capsid Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D005260:Female; D006801:Humans; D007114:Immunization; D007218:Industrial Microbiology / Q000295:instrumentation / Q000379:methods*; D051379:Mice; D008807:Mice, Inbred BALB C; D030361:Papillomavirus Infections / Q000821:virology*; D053918:Papillomavirus Vaccines / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D018411:Spodoptera
chemical_list: D036022:Capsid Proteins; D053918:Papillomavirus Vaccines; C091560:L1 protein, Human papillomavirus type 6
keywords: Bioreactor; Hollow fiber membrane; Human papillomavirus type-6; Sf9 cells; Virus-like particle