Microcarrier culture of COS-1 cells producing Japanese encephalitis and dengue virus serotype 4 recombinant virus-like particles.
Two stably transfected COS-1 cell lines that secrete recombinant Japanese encephalitis and dengue virus serotype 4 virus-like particles (VLPs) have been adapted to grow on Cytodex 3 microcarriers in an orbital shaker flask platform. The VLPs are used as antigens in diagnostic enzyme-linked immunosorbent assays to detect anti-arboviral IgM and IgG antibodies in human serum samples. Converting from a stationary flask batch system to a microcarrier fed-batch system has led to increases in antigen concentration while decreasing costs by reducing the amount of cell culture medium, disposables, and labor. The cell culture longevity was increased by 48 days in this optimized system, which may be due to a continual supply of nutrients resulting in prolonged survival of the cells on the microcarrier surface. An initial trial using a serum-free medium with this cell line was promising and may lead to reductions in cost, while reducing the variability between batches introduced by fetal bovine serum.
Velez
Jason O
JO
Diagnostic Reference Laboratory, Arbovirus Diseases Branch, Division of Vector-Borne Infectious Diseases, Centers for Disease Control and Prevention, 3150 Rampart Road., Fort Collins, CO 80521, United States.
Russell
Brandy J
BJ
Diagnostic Reference Laboratory, Arbovirus Diseases Branch, Division of Vector-Borne Infectious Diseases, Centers for Disease Control and Prevention, 3150 Rampart Road., Fort Collins, CO 80521, United States.
Hughes
Holly R
HR
Diagnostic Reference Laboratory, Arbovirus Diseases Branch, Division of Vector-Borne Infectious Diseases, Centers for Disease Control and Prevention, 3150 Rampart Road., Fort Collins, CO 80521, United States.
Chang
Gwong-Jen J
GJ
Arbovirus Diseases Branch, Division of Vector-Borne Infectious Diseases, Centers for Disease Control and Prevention, 3150 Rampart Road., Fort Collins, CO 80521, United States.
Johnson
Barbara W
BW
Diagnostic Reference Laboratory, Arbovirus Diseases Branch, Division of Vector-Borne Infectious Diseases, Centers for Disease Control and Prevention, 3150 Rampart Road., Fort Collins, CO 80521, United States.
D000818:Animals; D000956:Antigens, Viral / Q000032:analysis; D019556:COS Cells; D002522:Chlorocebus aethiops; D003716:Dengue Virus / Q000145:classification / Q000302:isolation & purification / Q000502:physiology*; D004664:Encephalitis Virus, Japanese / Q000302:isolation & purification / Q000502:physiology*; D012703:Serotyping; D014779:Virus Replication
D000956:Antigens, Viral