针对 SARS-COV-2 设计的多表位疫苗的表达、纯化和抗体反应评估以及抗体-免疫原复合物模拟。

IF 1 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Protein and Peptide Letters Pub Date : 2024-08-19 DOI:10.2174/0109298665320319240809095727
Ghadir A Jamal, Ehsan Jahangirian, Hossein Tarrahimofrad
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引用次数: 0

摘要

背景:COVID-19 疾病的传播是 SARS-CoV2 病毒感染的结果。冠状病毒中的四种关键蛋白:钉螺蛋白[S]、膜蛋白[M]、核壳蛋白[N]和包膜蛋白[E]在很大程度上被认为是冠状病毒的关键蛋白:本研究旨在表达多表位免疫原构建体的重组蛋白,并评估先前设计的作为 SARS-Cov-2 候选免疫原的多表位疫苗的免疫原性:将质粒 pET26b 转入表达宿主大肠杆菌 BL21 [DE3],在 IPTG 诱导下表达重组蛋白。重组蛋白经 Ni-NTA 柱亲和层析纯化,并用 Western 印迹法确认。最后,用重组蛋白分三次免疫小鼠。然后,通过对接和分子动力学模拟评估了疫苗三维结构与人类中和抗体三维结构[7BWJ和7K8N]抗体的相互作用:结果:优化基因的密码子兼容性指数为 0.96。在大肠杆菌宿主中表达 SARS-Cov-2 疫苗的重组蛋白,可产生重量约为 70 kDa、浓度为 0.7 mg/ml 的重组蛋白。用 SARS-Cov-2 疫苗重组蛋白免疫小鼠,可诱导 IgG 血清抗体反应。统计分析表明,抗体滴度与对照样品相比有显著差异,稀释至 1/256000时抗体滴度可接受。通过分子动力学模拟疫苗与人类抗体的结合,结果表明均方根偏差(RMSD)、均方根波动(RMSF)、回旋半径、H 键以及分子力学泊松-波尔兹曼表面积(MM/PBSA)分析的范德华能和静电都具有稳定的相互作用:结论:该重组蛋白可作为候选免疫原,在未来的研究中用于开发 SARS-CoV2 疫苗。
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Expression, Purification, and Evaluation of Antibody Responses and Antibody-Immunogen Complex Simulation of a Designed Multi-Epitope Vaccine against SARS-COV-2.

Background: The spread of the COVID-19 disease is the result of an infection caused by the SARS-CoV2 virus. Four crucial proteins, spike [S], membrane [M], nucleocapsid [N], and envelope [E] in coronaviruses have been considered to a large extent.

Objective: This research aimed to express the recombinant protein of a multiepitope immunogen construct and evaluate the immunogenicity of the multiepitope vaccine that was previously designed as a candidate immunogenic against SARS-Cov-2.

Materials and methods: Plasmid pET26b was transferred to the expression host E. coli BL21 [DE3] and the recombinant protein was expressed with IPTG induction. The recombinant protein was purified by Ni-NTA column affinity chromatography, and western blotting was used to confirm it. Finally, mice were immunized with recombinant protein in three doses. Then, the interaction of the 3D structure of the vaccine with the human neutralizing antibodies3D structures [7BWJ and 7K8N] antibody was evaluated by docking and molecular dynamics simulation.

Results: The optimized gene had a codon compatibility index of 0.96. The expression of the recombinant protein of the SARS-Cov-2 vaccine in an E. coli host led to the production of the recombinant protein with a weight of about 70 kDa with a concentration of 0.7 mg/ml. Immunization of mice with recombinant protein of SARS-Cov-2 vaccine-induced IgG serum antibody response. Statistical analysis showed that the antibody titer in comparison with the control sample has a significant difference, and the antibody titer was acceptable up to 1/256000 dilution. The simulation of vaccine binding with human antibodies by molecular dynamics showed that Root Mean Square Deviation [RMSD], Root Mean Square Fluctuation [RMSF], Radius of Gyration, and H-bond as well as van der Waals energies and electrostatic of Molecular mechanics Poisson- Boltzmann surface area [MM/PBSA] analysis have stable interaction.

Conclusion: This recombinant protein can probably be used as an immunogen candidate for the development of vaccines against SARS-CoV2 in future research.

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来源期刊
Protein and Peptide Letters
Protein and Peptide Letters 生物-生化与分子生物学
CiteScore
2.90
自引率
0.00%
发文量
98
审稿时长
2 months
期刊介绍: Protein & Peptide Letters publishes letters, original research papers, mini-reviews and guest edited issues in all important aspects of protein and peptide research, including structural studies, advances in recombinant expression, function, synthesis, enzymology, immunology, molecular modeling, and drug design. Manuscripts must have a significant element of novelty, timeliness and urgency that merit rapid publication. Reports of crystallization and preliminary structure determination of biologically important proteins are considered only if they include significant new approaches or deal with proteins of immediate importance, and preliminary structure determinations of biologically important proteins. Purely theoretical/review papers should provide new insight into the principles of protein/peptide structure and function. Manuscripts describing computational work should include some experimental data to provide confirmation of the results of calculations. Protein & Peptide Letters focuses on: Structure Studies Advances in Recombinant Expression Drug Design Chemical Synthesis Function Pharmacology Enzymology Conformational Analysis Immunology Biotechnology Protein Engineering Protein Folding Sequencing Molecular Recognition Purification and Analysis
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