Author: Kim, Eun; Erdos, Geza; Huang, Shaohua; Kenniston, Thomas; Falo, Louis D.; Gambotto, Andrea
Title: Preventative Vaccines for Zika Virus Outbreak: Preliminary Evaluation Document date: 2016_10_3
ID: jzcyxjxt_17
Snippet: We generated recombinant E1/E3-deleted adenovirus serotype 5based vectors that encode for the human secretory signal peptide hidden Markov model (SP-HMM) followed by the codon-optimized extracellular portion of the ZIKV BeH815744 E gene fused to the T4 fibritin foldon trimerization domain (ZIKV-Efl). Moreover, the ZIKV-Efl antigen was engineered with a polyhistidine-tag and a Tobacco Etch Virus (TEV) protease cleavage sequence to facilitate downs.....
Document: We generated recombinant E1/E3-deleted adenovirus serotype 5based vectors that encode for the human secretory signal peptide hidden Markov model (SP-HMM) followed by the codon-optimized extracellular portion of the ZIKV BeH815744 E gene fused to the T4 fibritin foldon trimerization domain (ZIKV-Efl). Moreover, the ZIKV-Efl antigen was engineered with a polyhistidine-tag and a Tobacco Etch Virus (TEV) protease cleavage sequence to facilitate downstream purification (Fig. 1) . The replication-defective adenovirus 5, designated as Ad5.ZIKV-Efl, was generated by loxP homologous recombination as previously described (Kim et al., 2014; Hardy et al., 1997) . Recombinant ZIKV-rEfl protein was purified from the supernatant of a Ad5.ZIKV-Eflinfected HEK 293 cell line using His60 Ni Superflow Resin under native conditions and packaged as a subunit vaccine in an MNA (MNA-ZIKV-Efl).
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