Development and evaluation of candidate vaccine and antitoxin against botulinum neurotoxin serotype F

Yu, Yun-Zhou; Zhang, Shuming; Ma, Yao; Zhu, Heng-Qi; Wang, Wenbing; Du, Yinan; Zhou, Xiaowei; Wang, Shuang; Yu, Wei-Yuan; Huang, Pei-Tang; Sun, Zhiwei

doi:10.1016/j.clim.2010.07.005

Cited by 16 publications

(15 citation statements)

References 30 publications

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“…Purification and characterization of the recombinant Hc proteins of serotypes A, B and F have been completed, demonstrating good safety and efficacy against botulism. [23][24][25][26][27][28] In this study, we presented the production and evaluation of a recombinant non-His-tagged Hc subunit vaccine as a candidate subunit vaccine for human use against BoNT/E. The prokaryotic expression vector pTIG-Trx-EHc was successfully constructed.…”

Section: Discussionmentioning

confidence: 99%

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Development and evaluation of candidate subunit vaccine and novel antitoxin against botulinum neurotoxin serotype E

Shi¹,

Liu

Mao³

et al. 2019

Human Vaccines & Immunotherapeutics

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Botulinum neurotoxins (BoNTs) are among the most toxic proteins. Vaccination is an effective strategy to prevent botulism. To generate a vaccine suitable for human use, a recombinant non-His-tagged isoform of the Hc domain of botulinum neurotoxin serotype E (rEHc) was expressed in Escherichia coli and purified by sequential chromatography. The immunogenicity of rEHc was evaluated in mice and doseand time-dependent immune responses were observed in both antibody titers and protective potency. Then, the pilot-scale expression and purification of rEHc were performed, and its immunological activity was characterized. Our results showed rEHc has good immunogenicity and can elicit strong protective potency against botulinum neurotoxin serotype E (BoNT/E) in mice, indicating that rEHc is an effective botulism vaccine candidate. Further, we developed a novel antitoxin against BoNT/E by purifying F(abʹ) 2 from pepsin-digested serum IgG of rEHc-inoculated horses. The protective effect of the F(abʹ) 2 antitoxin was determined in vitro and in vivo. The results showed that our F(abʹ) 2 antitoxin can prevent botulism in BoNT/E-challenged mice and effectively alleviate the progression of paralysis caused by BoNT/E to achieve therapeutic effects. Therefore, our results provide valuable experimental data for the production of a novel antitoxin, which is a promising candidate for the treatment of BoNT/E-induced botulism. ARTICLE HISTORY

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Section: Discussionmentioning

confidence: 99%

“…Total equine IgG and F(abʹ) 2 were prepared as previously described. 27,33,34 Briefly, the hyperimmune sera were diluted with pyrogen-free distilled water (1:2 v/v) and adjusted to pH 3.0 with 1 N HCl. The IgG was digested with pepsin (6 U/ml) at 37°C for 30 min.…”

Section: Preparation Of Equine Igg and F(ab') 2 Fragmentsmentioning

confidence: 99%

Development and evaluation of candidate subunit vaccine and novel antitoxin against botulinum neurotoxin serotype E

Shi¹,

Liu

Mao³

et al. 2019

Human Vaccines & Immunotherapeutics

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“…4,5,7 Extensive studies aimed to produce recombinant Hc of BoNTs as candidate vaccine are performed. The expression and purification of these proteins using E. coli [17][18][19][20][21] and yeast [22][23][24][25] are the systems used most widely. As an alternative, the Hc domain of BoNT/A as a subunit vaccine antigen was expressed using a bi-cistronic baculovirus-Sf21 insect cell expression system.…”

Section: Discussionmentioning

confidence: 99%

Production and evaluation of a recombinant subunit vaccine against botulinum neurotoxin serotype B using a 293E expression system

Shi²,

Liu³

et al. 2014

Human Vaccines & Immunotherapeutics

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Keywords: Botulinum neurotoxin serotype B, Hc, expression, vaccine, binding activityAlthough 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.

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“…To easy the cloning of foreign genes and the purification of soluble proteins, we first add one multiple clone site sequence (GAA TTC TAA TGG GCA GCA GCC ATC ATC ATC ATC ATC ACA GCA GCG GCC TGG TGC CGC GCG GGA GCT CGT CGA CAA GCT TGG TAC CTC GAG) in pTIG-Trx by EcoR I and Xho I resulting in a pTIG-Trx-M expression vector. 33,34 The oligonucleotide primers for PCR amplification of the synthetic AHc-C gene (amino acids 1,088 through 1,296, ~25 kDa) using pGEM-AHc as a template were forward primer, F-Hc-CS (Sac I): 5'-CAT GGA GCT CTT ACG ACA ACC AGT CCA ATT CTG G-3', and reverse primer R-Hc-CX (Xho I): 5'-CAT GCT CGA GCA GTG GAC GTT CAC CCC AAC-3' (underlined sequence indicate restriction enzyme recognition sites). The PCR products were digested with Sac I and Xho I to excise the AHc-C DNA fragment which was subcloned into a pTIG-Trx-M expression vector (cut with the same restriction enzymes) resulting in recombinant plasmid pTIG-Trx-AHc-C that was confirmed by sequencing.…”

Section: Methodsmentioning

confidence: 99%

Binding activity and immunogenic characterization of recombinant C-terminal quarter and half of the heavy chain of botulinum neurotoxin serotype A

Yu¹,

Ma²,

Chen³

et al. 2011

Human Vaccines

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Development and evaluation of candidate vaccine and antitoxin against botulinum neurotoxin serotype F

Cited by 16 publications

References 30 publications

Development and evaluation of candidate subunit vaccine and novel antitoxin against botulinum neurotoxin serotype E

Development and evaluation of candidate subunit vaccine and novel antitoxin against botulinum neurotoxin serotype E

Production and evaluation of a recombinant subunit vaccine against botulinum neurotoxin serotype B using a 293E expression system

Binding activity and immunogenic characterization of recombinant C-terminal quarter and half of the heavy chain of botulinum neurotoxin serotype A

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