Time filter

Source Type

Markely L.R.A.,Cell Culture Development High Throughput Analytical GroupBiogen | Cheung L.,Cell Culture Development High Throughput Analytical GroupBiogen | Choi Y.J.,Cell Culture Development High Throughput Analytical GroupBiogen | Ryll T.,Cell Culture Development High Throughput Analytical GroupBiogen | And 9 more authors.
Biotechnology Progress | Year: 2015

The serum half-life, biological activity, and solubility of many recombinant glycoproteins depend on their sialylation. Monitoring glycoprotein sialylation during cell culture manufacturing is, therefore, critical to ensure product efficacy and safety. Here a high-throughput method for semi-quantitative fingerprinting of glycoprotein sialylation using capillary isoelectric focusing immunoassay on NanoPro (Protein Simple) platform was developed. The method was specific, sensitive, precise, and robust. It could analyze 2 μL of crude cell culture samples without protein purification, and could automatically analyze from 8 samples in 4 h to 96 samples in 14 h without analyst supervision. Furthermore, its capability to detect various changes in sialylation fingerprints during cell culture manufacturing process was indispensable to ensure process robustness and consistency. Moreover, the changes in the sialylation fingerprints analyzed by this method showed strong correlations with intact mass analysis using liquid chromatography and mass spectrometry. © 2015 American Institute of Chemical Engineers.

Matthews T.E.,Inc5000 Davis Driveresearch Triangle Park27709North Carolina | Berry B.N.,Cell Culture DevelopmentBiogen | Smelko J.,Inc5000 Davis Driveresearch Triangle Park27709North Carolina | Moretto J.,Inc5000 Davis Driveresearch Triangle Park27709North Carolina | And 2 more authors.
Biotechnology and Bioengineering | Year: 2016

Accumulation of lactate in mammalian cell culture often negatively impacts culture performance, impeding production of therapeutic proteins. Many efforts have been made to limit the accumulation of lactate in cell culture. Here, we describe a closed loop control scheme based on online spectroscopic measurements of glucose and lactate concentrations. A Raman spectroscopy probe was used to monitor a fed-batch mammalian cell culture and predict glucose and lactate concentrations via multivariate calibration using partial least squares regression (PLS). The PLS models had a root mean squared error of prediction (RMSEP) of 0.27g/L for glucose and 0.20g/L for lactate. All glucose feeding was controlled by the Raman PLS model predictions. Glucose was automatically fed when lactate levels were beneath a setpoint (either 4.0 or 2.5g/L) and glucose was below its own setpoint (0.5g/L). This control scheme was successful in maintaining lactate levels at an arbitrary setpoint throughout the culture, as compared to the eventual accumulate of lactate to 8.0g/L in the historical process. Automated control of lactate by restricted glucose feeding led to improvements in culture duration, viability, productivity, and robustness. Culture duration was extended from 11 to 13 days, and harvest titer increased 85% over the historical process. © 2016 Wiley Periodicals, Inc.

Discover hidden collaborations