Monitoring the progress of infection and recombinant protein production in insect cell cultures using intracellular ATP measurement.
Several monitoring methods used to predict viable cell density have been the subject of extensive studies, including oxygen uptake rate, carbon dioxide evolution rate, optical density, NADH-dependent fluorescence and relative permittivity measurement. We propose intracellular ATP determination by bioluminescence assay to monitor the progress of baculovirus infection and recombinant protein production in insect cell cultures. We found that the ATP content in viable cells increased after virus addition. The increase in the ATP level was observed until the maximum recombinant protein accumulation was reached. At maximum product yield, the specific ATP content significantly decreased. Results obtained in both batch and fed-batch cultures demonstrated that the specific ATP level could be considered as a good indicator of recombinant protein productivity. Monitoring the cellular ATP content after viral infection makes it possible to define the optimum time for product harvest. The main advantage of applying the ATP assay as an index of the progress of infection and recombinant protein synthesis is its short time and sensitivity.
Olejnik
A M
AM
Department of Biotechnology and Food Microbiology, Agricultural University in Poznan, Wojska Polskiego 48 Str., 60-627 Poznan, Poland. aolejnik@owl.au.poznan.pl
Czaczyk
K
K
Marecik
R
R
Grajek
W
W
Jankowski
T
T
D000255:Adenosine Triphosphate / Q000378:metabolism*; D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001709:Biotechnology / Q000379:methods; D002455:Cell Division; D002478:Cells, Cultured; D005822:Genetic Vectors; D007915:Lepidoptera / Q000166:cytology; D008991:Monitoring, Physiologic / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D019076:Transgenes / Q000502:physiology*
D011994:Recombinant Proteins; D000255:Adenosine Triphosphate