Vaccine Analytical Research and Development, Merck & Co., Inc., West Point, PA, USA.
Vaccine Process Research and Development, Merck & Co., Inc., West Point, PA, USA.
Vaccine. 2022 Sep 2;40(37):5529-5536. doi: 10.1016/j.vaccine.2022.08.006. Epub 2022 Aug 9.
Vaccine development is a complex process, starting with selection of a promising immunogen in the discovery phase, followed by process development in the preclinical phase, and later by clinical trials in tandem with process improvements and scale up. A large suite of analytical techniques is required to gain understanding of the vaccine candidate so that a relevant immunogen is selected and subsequently manufactured consistently throughout the lifespan of the product. For viral vaccines, successful immunogen production is contingent on its maintained antigenicity and/or infectivity, as well as the ability to characterize these qualities within the context of the process, formulation, and clinical performance. In this report we show the utility of flow virometry during preclinical development of a Covid 19 vaccine candidate based on SARS-CoV-2 spike (S) protein expressed on vesicular stomatitis virus (VSV). Using a panel of monoclonal antibodies, we were able to detect the S protein on the surface of the recombinant VSV virus, monitor the expression levels, detect differences in the antigen based on S protein sequence and after virus inactivation, and monitor S protein stability. Collectively, flow virometry provided important data that helped to guide preclinical development of this vaccine candidate.
疫苗开发是一个复杂的过程,从发现阶段选择有前途的免疫原开始,然后在临床前阶段进行工艺开发,随后与工艺改进和放大并行进行临床试验。需要一整套分析技术来了解疫苗候选物,以便选择相关的免疫原,并在产品的整个生命周期内持续一致地制造。对于病毒疫苗,成功的免疫原生产取决于其保持的抗原性和/或感染力,以及在工艺、配方和临床性能背景下对这些特性进行表征的能力。在本报告中,我们展示了在基于 SARS-CoV-2 刺突(S)蛋白表达的新型冠状病毒(COVID-19)疫苗候选物的临床前开发过程中使用流式病毒测量的效用。使用一组单克隆抗体,我们能够检测到重组水疱性口炎病毒(VSV)病毒表面的 S 蛋白,监测表达水平,根据 S 蛋白序列和病毒失活后检测抗原差异,并监测 S 蛋白稳定性。总之,流式病毒测量提供了重要的数据,有助于指导该疫苗候选物的临床前开发。