Development of an automated, high-throughput SARS-CoV-2 neutralization assay based on a pseudotyped virus using a vesicular stomatitis virus (VSV) vector.
Ziteng LiangXi WuJiajing WuShuo LiuJincheng TongTao LiYuanling YuLi ZhangChenyan ZhaoQiong LuHaiyang QinJianhui NieWeijin HuangYou-Chun WangPublished in: Emerging microbes & infections (2023)
The global outbreak of COVID-19 has caused a severe threat to human health; therefore, simple, high-throughput neutralization assays are desirable for developing vaccines and drugs against COVID-19. In this study, a high-titer SARS-CoV-2 pseudovirus was successfully packaged by truncating the C-terminus of the SARS-CoV-2 spike protein by 21 amino acids and infecting 293T cells that had been stably transfected with the angiotensin-converting enzyme 2 (ACE2) receptor and furin (named AF cells), to establish a simple, high-throughput, and automated 384-well plate neutralization assay. The method was optimized for cell amount, virus inoculation, incubation time, and detection time. The automated assay showed good sensitivity, accuracy, reproducibility, Z' factor, and a good correlation with the live virus neutralization assay. The high-throughput approach would make it available for the SARS-CoV-2 neutralization test in large-scale clinical trials and seroepidemiological surveys which would aid the accelerated vaccine development and evaluation.
Keyphrases
- high throughput
- sars cov
- single cell
- angiotensin converting enzyme
- respiratory syndrome coronavirus
- human health
- clinical trial
- angiotensin ii
- amino acid
- induced apoptosis
- coronavirus disease
- atrial fibrillation
- stem cells
- randomized controlled trial
- disease virus
- machine learning
- binding protein
- climate change
- oxidative stress
- cell therapy
- cross sectional
- deep learning
- study protocol
- endoplasmic reticulum stress
- drug induced
- loop mediated isothermal amplification