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  • Application of the Bradford Assay for Cell Lysis Quantification: Residual Protein Content in Cell Culture Supernatants.

Application of the Bradford Assay for Cell Lysis Quantification: Residual Protein Content in Cell Culture Supernatants.

Biotechnology journal (2019-04-16)
Bernhard Sissolak, Christian Zabik, Natasa Saric, Wolfgang Sommeregger, Karola Vorauer-Uhl, Gerald Striedner
ABSTRACT

Frequently measured mammalian cell culture process indicators include viability and total cell concentration (TCC). Cell lysis, an additional important process characteristic that substantially contributes to the overall product purity profiles, is often not addressed in detail. In the present study, an inexpensive and simple application of the Bradford assay is developed to determine the residual protein content (RPC) in cell culture supernatants. The reliability and reproducibility of the method are tested in a long-term study and compared with lysis quantification via the DNA measurement. The results show that its performance is more robust and accurate over time and the respective concentration range. Additionally, both methods are used for cell lysis process monitoring in a recombinant Chinese hamster ovary fed-batch process. In the presented process, by applying the established assay, the lysis rate k DL is determined to be constant over time at 4.6 × 10 -4 lysed cell concentration (LCC) per TCC and time (LCC/TCC/h). In contrast, DNA data did not confirm the constant lysis rate due to variations of the content per cell during cultivation. Thus, information on the RPC can facilitate the determination of the optimal harvest time point with respect to purity and in improving process characterization.

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Sigma-Aldrich
Antifoam C Emulsion, aqueous-silicone emulsion