Synthesis of rotavirus-like particles in insect cells: comparative and quantitative analysis.
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.
Jiang
B
B
Wyeth-Lederle Vaccines and Pediatrics, 401 North Middletown Road, Pearl River, New York 10965, USA.
Barniak
V
V
Smith
R P
RP
Sharma
R
R
Corsaro
B
B
Hu
B
B
Madore
H P
HP
D000818:Animals; D000956:Antigens, Viral; D016367:Baculoviridae / Q000235:genetics; D002213:Capsid / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification; D036022:Capsid Proteins; D018929:Cell Culture Techniques / Q000379:methods; D002460:Cell Line; D016895:Culture Media, Serum-Free; D005822:Genetic Vectors; D007313:Insecta; D007700:Kinetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification; D012401:Rotavirus / Q000235:genetics*; D018411:Spodoptera; D013997:Time Factors; D014162:Transfection / Q000379:methods
D000956:Antigens, Viral; D036022:Capsid Proteins; D016895:Culture Media, Serum-Free; D011994:Recombinant Proteins; C442545:VP2 protein, Rotavirus; C061332:VP6 protein, Rotavirus; C050779:VP7 protein, Rotavirus