Abstract
Background Traditionally antibody producing cells like hybridoma cells sank into oblivion since other suspension cell lines have captured the biopharmaceutical production market. However, they are still of particular interest in academic and industrial diagnostic research. Hence, fast and sufficient antibody production is needed as proof of concept, for toxicology and in vivo studies. Although, hybridoma cultivation in fetal bovine serum (FBS) containing animal derived ingredients, like contaminating IgG, is undesirable and leads to difficulties in purification. When reducing the serum to a minimum other key components of the FBS have to be replaced. Therefore, human insulinlike growth factor (IGF) [1] and the surfactant Pluronic F68 were supplemented to improve overall cell performance and to reduce shear stress during shaking respectively employing Design of Experiment (DoE)[2]. Compared to the original basal medium an improvement in cell growth, viability and antibody titer was achieved. These optimized inoculum conditions were used for subsequent bioreactor fermentations. Furthermore, these conditions were used in order to test feeding strategies. For this purpose a fed-batch process with a double bolus feed was simulated in shake flasks with two different glucose feeding strategies with and without Hyclone Cell Boost 6 (CB6). Finally, the result from shake flasks could be verified and improved antibody yield was achieved in a controlled 2L fed-batch process.
Highlights
Antibody producing cells like hybridoma cells sank into oblivion since other suspension cell lines have captured the biopharmaceutical production market
Feeding strategy Cells were seeded with 3 × 105 cells/mL in 35 mL working volume in 125 mL shake flasks in optimized medium (DMEM, 4.5 g/L glucose, 2 mM stable glutamine, 6% fetal bovine serum (FBS), 100 μg/L insulinlike growth factor (IGF) and 0.2 g/L Pluronic)
Design of Experiment (DoE) approach A simple DoE approach with the three factors FBS, IGF and Pluronic led to improved hybridoma cultivation
Summary
Antibody producing cells like hybridoma cells sank into oblivion since other suspension cell lines have captured the biopharmaceutical production market. They are still of particular interest in academic and industrial diagnostic research. Compared to the original basal medium an improvement in cell growth, viability and antibody titer was achieved. These optimized inoculum conditions were used for subsequent bioreactor fermentations. These conditions were used in order to test feeding strategies For this purpose a fed-batch process with a double bolus feed was simulated in shake flasks with two different glucose feeding strategies - with and without Hyclone Cell Boost 6 (CB6). The result from shake flasks could be verified and improved antibody yield was achieved in a controlled 2L fed-batch process
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