Prediction of recombinant protein production in an insect cell-baculovirus system using a flow cytometric technique.
The baculovirus expression vector system (BEVS) utilising the Autographa californica nucleopolyhedrovirus (AcMNPV) is widely becoming the system of choice for the production of many recombinant protein products due to the high yields obtained. However, there is a need to develop a simple reliable on-line method to monitor the production of recombinant proteins that have no intrinsic reporter properties. Here we utilise flow cytometry to measure cell size, granularity and DNA content in a single step analysis and correlate these parameters with the production of the recombinant protein beta-galactosidase. Clear correlations between these parameters and productivity are made with forward and side scatter signals showing the highest correlation coefficients. Measuring these parameters does not require any processing of the cells from culture to analysis. These parameters can therefore be used successfully to predict the amount of recombinant protein product in a BEVS system on-line.
Sandhu
Kalbinder Singh
KS
School of Chemical and Bioprocess Engineering, University College Dublin, Belfield, Dublin 4, Ireland.
Naciri
Mariam
M
Al-Rubeai
Mohamed
M
D000818:Animals; D002460:Cell Line; D048429:Cell Size; D002470:Cell Survival; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D017924:Nucleopolyhedroviruses / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis*; D018411:Spodoptera; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D016895:Culture Media, Serum-Free; D011994:Recombinant Proteins; D001616:beta-Galactosidase