Abstract
Background Process optimization and control are essential to match biopharmaceutical market demands. Early-steps in the development of new processes require screening hundreds of transfected clones in order to select those with the best expression characteristics, and identifying optimal environmental conditions for these clones to grow and express the protein of interest. While some culture systems include in situ analysis of cell density, most often by optical density or capacitance measurements, the majority of them still rely on off-line, laborious and time-consuming protocols for recombinant protein analysis. Spectroscopic methods have been proposed in literature to support bioprocesses development, because they are non-invasive and provide data on multiple components present in the culture bulks, thereby increasing information on process performance [1]. Here, we developed a fluorescence-based method for real-time monitoring of viable cells and antibody titers in bioreactor cultures, and extended this strategy for high throughput analysis of cellular productivity in 96well plates.
Highlights
Process optimization and control are essential to match biopharmaceutical market demands
The supernatant samples were distributed into black 96well plates and the fluorescence maps were collected in a spectrofluorometer (Horiba Jobin Yvon) connected to a
The fluorescence analysis of the culture bulk was performed in situ, whereas for the 96-well plate application the cultures were performed in shake flasks and only the cell supernatant was analyzed in terms of fluorescence
Summary
Process optimization and control are essential to match biopharmaceutical market demands. Early-steps in the development of new processes require screening hundreds of transfected clones in order to select those with the best expression characteristics, and identifying optimal environmental conditions for these clones to grow and express the protein of interest. While some culture systems include in situ analysis of cell density, most often by optical density or capacitance measurements, the majority of them still rely on off-line, laborious and time-consuming protocols for recombinant protein analysis. Spectroscopic methods have been proposed in literature to support bioprocesses development, because they are non-invasive and provide data on multiple components present in the culture bulks, thereby increasing information on process performance [1]. We developed a fluorescence-based method for real-time monitoring of viable cells and antibody titers in bioreactor cultures, and extended this strategy for high throughput analysis of cellular productivity in 96well plates
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