伪型化提高了功能性SARS-CoV-2病毒样粒子 (VLP) 的产量,作为疫苗和治疗开发的工具
Andrew J Zak1, Trang Hoang1, Christine M Yee1
1Department of Chemical Engineering, University of Michigan, Ann Arbor, MI 48109, USA.
International journal of molecular sciences
|October 14, 2023
概括
这项研究通过伪型化增强了SARS-CoV-2病毒样粒子 (VLP) 的产生,显著增加了尖端产量和抗原密度. 这些改进的VLP对于开发疫苗和中和抗体测试对抗SARS-CoV-2变种非常有价值.
科学领域:
- 生物技术是生物技术.
- 病毒学 病毒学
- 疫苗开发 疫苗开发
背景情况:
- 类似病毒的粒子 (VLP) 对SARS-CoV-2疫苗和测试具有前景.
- 以前的VLP设计具有较低的尖峰产量,限制了它们的实用性.
- 量化VLP中的尖端蛋白质含量对于平台的可行性至关重要.
研究的目的:
- 为了改善SARS-CoV-2 VLP的产量和抗原密度.
- 为各种SARS-CoV-2变种开发功能VLP.
- 在测试中证明伪型VLP作为活病毒的替代品.
主要方法:
- 通过将其与流感HA的跨膜域融合,设计了SARS-CoV-2尖端蛋白.
- 在Sf9昆虫细胞中与M1蛋白共同表达的改性尖蛋白.
- 为不同的SARS-CoV-2变种 (阿尔法,贝塔,三角形,奥米克朗) 制作和特征化伪型VLP.
主要成果:
- 与原生设计 (~0.04-0.08 mg/L) 相比,伪造型显著增加了VLP峰值产量 (~0.4 mg/L).
- 达到更高的VLP尖峰抗原密度 (~96个单体/VLP),更接近本地病毒.
- 为结合ACE2的多个SARS-CoV-2变种生成功能伪型VLP.
- 在BSL-1 ELISA试验中证明了伪型VLP用于中和抗体检测.
结论:
- 伪造型比本地设计更有利于更高的SARS-CoV-2 VLP尖端产量.
- 伪型VLP在疫苗和治疗开发中作为活体SARS-CoV-2的有效替代品.
- 这种方法可以在安全的实验室条件下对SARS-CoV-2变种进行研究.
相关概念视频
02:50Conjugated Proteins
Simple proteins and protein complexes contain only amino acids. In contrast, many other proteins, called conjugated proteins, covalently bond with non-protein moieties.
Nucleoproteins are protein complexes that contain nucleic acids, categorized as deoxyribonucleoproteins (DNPs) or ribonucleoproteins (RNPs) respectively. The nucleosome is a typical example of a DNP where nuclear DNA is associated with histone proteins. The major antigen for the Covid-19 virus SARS-CoV is an RNP that is critical...
Nucleoproteins are protein complexes that contain nucleic acids, categorized as deoxyribonucleoproteins (DNPs) or ribonucleoproteins (RNPs) respectively. The nucleosome is a typical example of a DNP where nuclear DNA is associated with histone proteins. The major antigen for the Covid-19 virus SARS-CoV is an RNP that is critical...
02:28Leaky Scanning
During most eukaryotic translation processes, the small 40S ribosome subunit scans an mRNA from its 5' end until it encounters the first start AUG codon. The large 60S ribosomal subunit then joins the smaller one to initiate protein synthesis. The location of the translation initiation is largely determined by the nucleotides near the start codon as there may be multiple translation initiation sites present on the mRNA. Marilyn Kozak discovered that the sequence RCCAUGG (where R stands for...
01:23Vaccine Production
Vaccine production involves a sequence of upstream and downstream processes to generate a safe and effective immunological product. It begins with cultivating microorganisms, such as viruses or bacteria, to obtain antigenic material. For viral vaccines, mammalian host cells are grown in bioreactors and subsequently infected with the target virus. The virus replicates within the host cells, which are lysed to release viral particles. This lysate is then clarified through filtration or...
01:29Coronavirus
Coronaviruses, including the severe acute respiratory syndrome coronavirus (SARS-CoV), are enveloped viruses characterized by their single-stranded, positive-sense RNA genome and helical nucleocapsid structure. The hallmark of these viruses is their club-shaped spike (S) glycoproteins that protrude from the viral envelope, facilitating attachment to host cells. Typically, coronaviruses infect the upper respiratory tract, often causing mild or asymptomatic disease. However, certain strains like...