RT Journal Article
SR Electronic
T1 A common problem in the purification of calicivirus raised by molecular sieving: balance between purification and loss of viral particles
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.02.04.933390
DO 10.1101/2020.02.04.933390
A1 Zehui Qu
A1 Hongtao Kang
A1 Chenxi Cui
A1 Kaiwen Meng
A1 Xinzheng Zhang
A1 Liandong Qu
A1 Geng Meng
YR 2020
UL http://biorxiv.org/content/early/2020/02/04/2020.02.04.933390.abstract
AB Caliciviridae is a group of RNA viruses could lead to gastrointestinal disease in humans and acute oral or upper respiratory tract disease in felines, no efficient vaccine so far. The preparation of immunogen is a challenge to obtain a safe and efficient vaccine candidate. Molecular sieving, due to the advantage of simple and fast purifying, is considered as a very powerful tool in calicivirus purification; however, also introduce mechanical damage to the viron. In this study, we took Feline Calicivirus (FCV), a common natural pathogen in cats as well as a classic model for caliciviruses studies, as a research object. Here, with the help of the cryo-electron microscope (cryo-EM), we collected particle images in both conditions of with and without molecular sieving post-processing, the impact of molecular sieving on FCV particles was evaluated by building a flexibility evaluation system after 3D classification. The results indicated that the molecular sieving will impact the stability of P domains through increasing flexibility. In addition, we successfully built up a scoring system to describe this flexibility of FCV P-domains, which can be extended to assess the purification effect to the entire calicivirus.