Kim Soo-Hyun, Lim Kwang-Il
Department of Chemical and Biological Engineering, Sookmyung Women's University, Seoul 04310, Korea.
Mol Cells. 2017 May 31;40(5):339-345. doi: 10.14348/molcells.2017.0043. Epub 2017 May 2.
Retroviral and lentiviral vectors are mostly pseudotyped and often purified and concentrated via ultracentrifugation. In this study, we quantified and compared the stabilities of retroviral [murine leukemia virus (MLV)-based] and lentiviral [human immunodeficiency virus (HIV)-1-based] vectors pseudotyped with relatively mechanically stable envelope proteins, vesicular stomatitis virus glycoproteins (VSVGs), and the influenza virus WSN strain envelope proteins against ultracentrifugation. Lentiviral genomic and functional particles were more stable than the corresponding retroviral particles against ultracentrifugation when pseudotyped with VSVGs. However, both retroviral and lentiviral particles were unstable when pseudotyped with the influenza virus WSN strain envelope proteins. Therefore, the stabilities of pseudotyped retroviral and lentiviral vectors against ultracentrifugation process are a function of not only the type of envelope proteins, but also the type of viral internal core (MLV or HIV-1 core). In addition, the fraction of functional viral particles among genomic viral particles greatly varied at times during packaging, depending on the type of envelope proteins used for pseudotyping and the viral internal core.
逆转录病毒载体和慢病毒载体大多进行了假型化处理，并且常常通过超速离心进行纯化和浓缩。在本研究中，我们对用相对机械稳定的包膜蛋白、水泡性口炎病毒糖蛋白（VSVGs）以及流感病毒WSN株包膜蛋白进行假型化处理的逆转录病毒[基于鼠白血病病毒（MLV）]载体和慢病毒[基于人类免疫缺陷病毒（HIV）-1]载体在超速离心过程中的稳定性进行了定量和比较。当用VSVGs进行假型化处理时，慢病毒基因组和功能颗粒在超速离心过程中比相应的逆转录病毒颗粒更稳定。然而，当用流感病毒WSN株包膜蛋白进行假型化处理时，逆转录病毒颗粒和慢病毒颗粒均不稳定。因此，假型化逆转录病毒载体和慢病毒载体在超速离心过程中的稳定性不仅取决于包膜蛋白的类型，还取决于病毒内部核心（MLV或HIV-1核心）的类型。此外，在包装过程中，基因组病毒颗粒中功能病毒颗粒的比例有时会因用于假型化的包膜蛋白类型和病毒内部核心的不同而有很大差异。
