title: Production and evaluation of a recombinant subunit vaccine against botulinum neurotoxin serotype B using a 293E expression system.
abstract: Although Escherichia coli and yeast were commonly used to express recombinant Hc of botulinum neurotoxins, as an alternative, in current study, a 293E expression system was used to express the Hc of botulinum neurotoxin serotype B (BHc) as soluble recombinant protein for experimental vaccine evaluation. Our results demonstrated that the 293E expression system could produce high level of recombinant secreted BHc protein, which was immunorecognized specifically by anti-botulinum neurotoxin serotype B (BoNT/B) sera and showed ganglioside binding activities. The serological response and efficacy of recombinant BHc formulated with aluminum hydroxide adjuvant were evaluated in mice. Immunization with Alhydrogel-formulated BHc subunit vaccine afforded the effective protection against BoNT/B challenge. A frequency- and dose-dependent effect to immunization with BHc subunit vaccine was observed and the ELISA antibody titers correlated well with neutralizing antibody titers and protection. And a solid-phase assay showed that the neutralizing antibodies from the BHc-immunized mice inhibited the binding of BHc to the ganglioside GT1b. Our results also show that the plasmid pABE293SBHc derived of the 293E expression system as DNA vaccine is capable of inducing stronger humoral response and protective efficacy against BoNT/B than the pVAX1SBHc. In summary, immunization with the 293E-expressed BHc protein generates effective immune protection against BoNT/B as E. coli or yeast-expressed BHc, so the efficient expression of botulinum Hc protein for experimental vaccine can be prepared using the 293E expression system.
authors.affiliation: a Beijing Institute of Biotechnology ; Beijing , PR China.
authors.forename: YunZhou
authors.initials: Y
authors.lastname: Yu
authors.forename: DanYang
authors.initials: D
authors.lastname: Shi
authors.forename: Si
authors.initials: S
authors.lastname: Liu
authors.forename: Zheng-Wei
authors.initials: ZW
authors.lastname: Gong
authors.forename: Shuang
authors.initials: S
authors.lastname: Wang
authors.forename: ZhiWei
authors.initials: Z
authors.lastname: Sun
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage; D000536:Aluminum Hydroxide / Q000008:administration & dosage; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D057134:Antibodies, Neutralizing / Q000097:blood; D000992:Antitoxins / Q000097:blood; D001428:Bacterial Vaccines / Q000008:administration & dosage / Q000276:immunology* / Q000302:isolation & purification; D019274:Botulinum Toxins, Type A / Q000276:immunology*; D001906:Botulism / Q000276:immunology / Q000517:prevention & control*; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D019444:Vaccines, DNA / Q000008:administration & dosage / Q000276:immunology / Q000302:isolation & purification; D022223:Vaccines, Subunit / Q000008:administration & dosage / Q000276:immunology / Q000302:isolation & purification; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000276:immunology / Q000302:isolation & purification
chemical_list: D000276:Adjuvants, Immunologic; D000907:Antibodies, Bacterial; D057134:Antibodies, Neutralizing; D000992:Antitoxins; D001428:Bacterial Vaccines; D019444:Vaccines, DNA; D022223:Vaccines, Subunit; D014614:Vaccines, Synthetic; C096323:rimabotulinumtoxinB; D000536:Aluminum Hydroxide; D019274:Botulinum Toxins, Type A
keywords: Botulinum neurotoxin serotype B; Hc; binding activity; expression; vaccine