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[Preparation and immungenicity of recombinant protein containing intramolecular adjuvant in SARS-CoV-2 RBD domain].
Jiang, Jingwen; Wang, Yunlong; Li, Yulin; Wang, Jichuang; Zhang, Yiqing; Wang, Xudong; Wang, Xiaojun; Zhang, Heng.
  • Jiang J; School of Life Sciences, Zhengzhou University, Zhengzhou 450001, Henan, China.
  • Wang Y; Henan Bioengineering Technology Research Center, Zhengzhou 450010, Henan, China.
  • Li Y; Department of Bioengineering, Zhengzhou Technical College, Zhengzhou 450010, Henan, China.
  • Wang J; Henan Bioengineering Technology Research Center, Zhengzhou 450010, Henan, China.
  • Zhang Y; Henan Bioengineering Technology Research Center, Zhengzhou 450010, Henan, China.
  • Wang X; Henan Bioengineering Technology Research Center, Zhengzhou 450010, Henan, China.
  • Wang X; Department of Bioengineering, Zhengzhou Technical College, Zhengzhou 450010, Henan, China.
  • Zhang H; Henan Bioengineering Technology Research Center, Zhengzhou 450010, Henan, China.
Sheng Wu Gong Cheng Xue Bao ; 38(9): 3353-3362, 2022 Sep 25.
Article in Chinese | MEDLINE | ID: covidwho-2264708
ABSTRACT
A fusion protein containing a tetanus toxin peptide, a tuftsin peptide and a SARS-CoV-2S protein receptor-binding domain (RBD) was prepared to investigate the effect of intramolecular adjuvant on humoral and cellular immunity of RBD protein. The tetanus toxin peptide, tuftsin peptide and S protein RBD region were connected by a flexible polypeptide, and a recombinant vector was constructed after codon optimization. The recombinant S-TT-tuftsin protein was prepared by prokaryotic expression and purification. BALB/c mice were immunized after mixed with aluminum adjuvant, and the humoral and cellular immune effects were evaluated. The recombinant S-TT-tuftsin protein was expressed as an inclusion body, and was purified by ion exchange chromatography and renaturated by gradient dialysis. The renaturated protein was identified by Dot blotting and reacted with serum of descendants immunized with SARS-CoV-2 subunit vaccine. The results showed that the antibody level reached a plateau after 35 days of immunization, and the serum antibody ELISA titer of mice immunized with recombinant protein containing intramolecular adjuvant was up to 166 240, which was significantly higher than that of mice immunized with S-RBD protein (P < 0.05). At the same time, the recombinant protein containing intramolecular adjuvant stimulated mice to produce a stronger lymphocyte proliferation ability. The stimulation index was 4.71±0.15, which was significantly different from that of the S-RBD protein (1.83±0.09) (P < 0.000 1). Intramolecular adjuvant tetanus toxin peptide and tuftsin peptide significantly enhanced the humoral and cellular immune effect of the SARS-CoV-2 S protein RBD domain, which provideda theoretical basis for the development of subunit vaccines for SARS-CoV-2 and other viruses.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Tuftsin / Viral Vaccines / COVID-19 Type of study: Experimental Studies Topics: Vaccines Limits: Animals / Humans Language: Chinese Journal: Sheng Wu Gong Cheng Xue Bao Journal subject: Biotechnology Year: 2022 Document Type: Article Affiliation country: J.cjb.210883

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Tuftsin / Viral Vaccines / COVID-19 Type of study: Experimental Studies Topics: Vaccines Limits: Animals / Humans Language: Chinese Journal: Sheng Wu Gong Cheng Xue Bao Journal subject: Biotechnology Year: 2022 Document Type: Article Affiliation country: J.cjb.210883