Abstract:
SARS-CoV-2, the causative agent of COVID-19, has imposed a major public health threat, which needs effective therapeutics and vaccination strategies. Several potential candidate vaccines being rapidly developed are in clinical evaluation and recombinant vaccine has gained much attention thanks to its potential for greater response predictability, improved efficacy, rapid development and reduced side effects. Recombinant vaccines are designed and manufactured using bacterial, yeast cells or mammalian cells. A small piece of DNA is taken from the virus or bacterium against which we want to protect and inserted into the manufacturing cells. Due to the extremely complex heterogeneity of SARS-CoV-2 recombinant vaccine, single technology platform cannot achieve thorough and accurate characterization of such difficult proteins so integrating comprehensive technologies is essential. This study illustrates an innovative workflow employing multiple separation techniques tandem high-resolution mass spectrometry for comprehensive and in-depth characterization of SARS-CoV-2 recombinant vaccine, including ultra-high performance liquid chromatography (UHPLC), ion exchange chromatography (IEX) and imaged capillary isoelectric focusing (icIEF). The integrated methodology focuses on the importance of cutting-edge icIEF-MS online coupling and icIEF fractionation applied to revealing the heterogeneity secret of SARS-CoV-2 recombinant vaccine.
摘要:
SARS-CoV-2是COVID-19的病原体,对公共卫生造成了重大威胁,这需要有效的治疗方法和疫苗接种策略。正在快速开发的几种潜在候选疫苗正在临床评估中,重组疫苗因其具有更大的反应可预测性而获得了广泛关注。提高疗效,快速发展和减少副作用。重组疫苗是使用细菌设计和制造的,酵母细胞或哺乳动物细胞。从我们想要保护的病毒或细菌中取出一小块DNA,并插入到制造细胞中。由于SARS-CoV-2重组疫苗具有极其复杂的异质性,单一的技术平台无法实现对这些困难蛋白质的彻底和准确的表征,因此整合综合技术至关重要。这项研究说明了一种创新的工作流程,采用多种分离技术串联高分辨率质谱来全面深入地表征SARS-CoV-2重组疫苗,包括超高效液相色谱(UHPLC),离子交换色谱(IEX)和成像毛细管等电聚焦(ICIEF)。综合方法侧重于尖端的icIEF-MS在线偶联和icIEF分级分离对揭示SARS-CoV-2重组疫苗异质性秘密的重要性。