Combinative workflow for mRNA vaccine development

IF 2.3 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochemistry and Biophysics Reports Pub Date : 2024-06-28 DOI:10.1016/j.bbrep.2024.101766

Renuka Khanzode , Garima Soni , Shalini Srivastava, Sharad Pawar, Rucha Wadapurkar, Ajay Singh

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引用次数: 0

Abstract

Recently, mRNA has gained a lot of attention in the field of vaccines, gene therapy, and protein replacement therapies. Herein, we are demonstrating a comprehensive approach to designing, cloning, and characterizing an antigenic cassette for the development of mRNA vaccine for COVID-19. The gene encoding the antigenic spike protein of the SARS-CoV-2 Omicron variant (B.1.1.529) was designed using the databases, characterized by in-silico tools, and assembled using overlapping oligonucleotide-based assembly by PCR. Next, the gene was cloned, mRNA was synthesized, and characterized using orthogonal approaches (Capillary electrophoresis, Sanger DNA sequencing, Next-generation sequencing, HPLC, qPCR, etc.). Furthermore, the antigen expression was monitored in-vitro using an animal cell model by western blot, flow cytometer, and surface plasmon resonance. The demonstrated approach has also been followed for developing the mRNA vaccines for various other indications such as Malaria, Herpes, Dengue, HPV, etc.

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mRNA 疫苗开发的组合工作流程

最近，mRNA 在疫苗、基因治疗和蛋白质替代疗法领域获得了广泛关注。在此，我们展示了一种用于开发 COVID-19 mRNA 疫苗的抗原盒的设计、克隆和表征的综合方法。我们利用数据库设计了编码 SARS-CoV-2 Omicron 变体（B.1.1.529）的抗原尖峰蛋白的基因，利用体内工具对其进行了表征，并利用基于寡核苷酸的重叠聚合酶链反应组装了该基因。接着，克隆了基因，合成了 mRNA，并使用正交方法（毛细管电泳、Sanger DNA 测序、下一代测序、HPLC、qPCR 等）对其进行了表征。此外，还通过 Western 印迹、流式细胞仪和表面等离子体共振等方法，利用动物细胞模型对抗原表达进行了体外监测。在开发用于疟疾、疱疹、登革热、人乳头瘤病毒等其他各种适应症的 mRNA 疫苗时，也采用了所展示的方法。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biochemistry and Biophysics Reports Biochemistry, Genetics and Molecular Biology-Biophysics

CiteScore

4.60

自引率

0.00%

发文量

191

审稿时长

59 days

期刊介绍： Open access, online only, peer-reviewed international journal in the Life Sciences, established in 2014 Biochemistry and Biophysics Reports (BB Reports) publishes original research in all aspects of Biochemistry, Biophysics and related areas like Molecular and Cell Biology. BB Reports welcomes solid though more preliminary, descriptive and small scale results if they have the potential to stimulate and/or contribute to future research, leading to new insights or hypothesis. Primary criteria for acceptance is that the work is original, scientifically and technically sound and provides valuable knowledge to life sciences research. We strongly believe all results deserve to be published and documented for the advancement of science. BB Reports specifically appreciates receiving reports on: Negative results, Replication studies, Reanalysis of previous datasets.