Quantification of cell lysis during CHO bioprocesses: Impact on cell count, growth kinetics and productivity.

摘要: High cell densities and high viability are critical quality attributes for mammalian bioprocesses. Determination of living dead numbers is nowadays routinely performed by automated image-based analyzers or flow cytometry. However, complete lysis cells usually neglected these devices. We present a novel method robust quantification lysed populations over the course CHO bioprocess. The release lactate dehydrogenase (LDH) double stranded genomic DNA in culture supernatants were used as markers lysis. considered degradation both cultivation time, which significantly increased amount released LDH DNA. For correct estimation fractions, time was considered, where redundancy allowed data reconciliation. Calculating number subject to lysis, we could show that this fraction makes up much 30% total produced biomass not described measurements analyzers. Finally, demonstrate disregarding heavily affects calculation yields growth rates increasing levels related decreased productivity.