Development of a simple and high-yielding fed-batch process for the production of influenza vaccines.
A robust and reliable GMP-compatible fed-batch process was successfully developed for the production of recombinant hemagglutinin (rHA) proteins by expresSF cells. The feeding solution, feeding strategy as well as the cell density at infection were optimized to maximize the final rHA production yields without affecting the existing rHA recovery protocol and downstream process. A simple and stable feeding solution was formulated and a rational feeding regimen designed to yield, depending on the rHA baculovirus used, between 2- and 3-fold enhancements in volumetric rHA production with increased specific productivity compared to the batch culture. Recombinant HA from fed-batch cultures could be simply recovered following cell lysis and purified through chromatographic steps. Overall, the increased rHA yield was maintained throughout the whole process. The performance, reproducibility and scalability of the fed-batch process was successfully demonstrated in 12 bioreactor runs of 2- and 10-L working volume using five different rHA encoding baculoviruses.
Meghrous
Jamal
J
Protein Sciences Corporation, 1000 Research Parkway, Meriden, CT 06450, United States.
Mahmoud
Wafaa
W
Jacob
Danielle
D
Chubet
Rick
R
Cox
Manon
M
Kamen
Amine A
AA
D000818:Animals; D002460:Cell Line; D006388:Hemagglutinins / Q000235:genetics / Q000378:metabolism; D007252:Influenza Vaccines / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D018411:Spodoptera
D006388:Hemagglutinins; D007252:Influenza Vaccines; D011994:Recombinant Proteins