Synthesis of rotavirus-like particles in insect cells: Comparative and quantitative analysis

Abstract

When the three major structural proteins, VP2, VP6, and VP7, of rotavirus are co‐expressed in insect cells infected with recombinant baculoviruses, they self‐assemble into triple‐layered virus‐like particles (VLPs) that are similar in morphology to native rotavirus. In order to establish the most favorable conditions for the synthesis of rotavirus VLPs, we have compared the kinetics of 2/6/7‐VLP synthesis in two different insect cell lines: High Five cells propagated in Excell 405 medium and Spodoptera frugiperda 9 cells in Excell 400 medium. The majority of VLPs produced in both cell lines were released into the culture medium, and these released VLPs were predominantly triple‐layered and were found to be stable for the period of six or seven days examined. The optimal synthesis of VLPs depended upon the cell line and the culture medium used as well as the time of harvesting infected cell cultures. The highest yield of VLPs was obtained from High Five cultures in the late phase of infection when the yield was at least 5‐fold higher than that from S. frugiperda 9 cultures on a per cell basis. Our results demonstrate the usefulness of High Five cells for the production of VLPs as potential rotavirus subunit vaccines. © 1998 John Wiley & Sons, Inc. Biotechnol Bioeng 60: 369–374, 1998.

Interaction of Hepatitis C Virus-Like Particles and Cells: a Model System for Studying Viral Binding and Entry

“…In the absence of purified HCV particles, the baculovirus-derived HCV-LP (5) has provided a surrogate model for this purpose. Generation of virus-like particles by using a recombinant baculovirus expression system has been successfully reported for many viruses such as papillomavirus (30), rotavirus (23), and JC polyomavirus (19). These virus-like particles have been demonstrated to be a valuable tool for investigating virus-cell interaction of papillomavirus (10) and for identifying the candidate receptor for Norwalk virus (46).…”

Rotavirus VP6 preparations as a non-replicating vaccine candidates

“…It has been shown by several studies that the structural proteins (VP2, VP6, VP7, and VP4) of RV can be expressed in different prokaryotic and eukaryotic expression systems and form SLPs (VP2), DLPs (VP2 and VP6), and TLPs (VP2, VP6, and VP7 or VP2, VP6, VP7, and VP4) [132][133][134][135][136][137][138][139][140][141][142][143][144][145][146][147][148]. Furthermore, expressing VP6 alone in vitro [14,29] can form spherical, tubular, and trimer structures similar to VP6 purified from the RVs [31].…”

Rotavirus-like particles primary recovery from insect cells in aqueous two-phase systems☆

“…An expression system that has been proven to be especially suitable for the production of VLP is the insect cellbaculovirus expression vector system (IC-BEVS) [8]. Although the production of dlRLP has been studied before, the existing reports addressed mainly the upstream part of the process [1,7,9,10]. In general, purification of VLP is a complex task.…”