Application of Roche Cedex Bio Biological Process Analyzer in Biotechnology Production Process Monitoring
D. Druhmann, S. Reinhard, F. Schwarz, C. Schaaf, K. Greisl, TL Nötzel
An essential element in the development and control of biological processes for the industrial production of recombinant proteins is the provision of fast, accurate and reliable process data. Accurate monitoring of matrices (nutrients) and metabolites in animal and bacterial cell cultures is essential to avoid undernutrition during fermentation or accumulation of toxic metabolic end products. Uncontrolled metabolites can adversely affect cell growth and survival as well as protein quality and yield. Therefore, accurate tracking of the fermentation process ensures reproducibility and is key to optimizing process development and validation.
Table: Comparison of detection ranges |
Typical parameters tested include glucose, lactic acid, glutamic acid, glutamine, ammonia, sodium and potassium. Currently, multi-parameter analyzer systems for detecting matrices and metabolites employ enzyme membrane biosensors and ion selective electrodes. The main disadvantage of these instruments is that the accuracy of the enzyme membrane decreases over time, the material cost is high, the test results are non-linear, and the overall sensitivity and accuracy are low.
Roche Cedex Bio Biological Process Analyzer
 | Figure 1: Accuracy and linear comparison of blood glucose, accuracy and linear comparison of lactic acid, accuracy and linear comparison of glutamine |
The Cedex Bio Bioprocess Analyzer and the recently introduced Cedex Bio HT Bioprocess Analyzer can be used to enhance process monitoring during fermentation. The Cedex Bio HT Bioprocess Analyzer is designed for high throughput testing of process development with up to 320 tests per hour. The results of the Cedex Bio HT Bioprocess Analyzer are identical to those obtained with the Cedex Bio Bioprocess Analyzer. This technology uses Roche's proven instrument platform, which significantly improves sensitivity and reproducibility over its extended detection range compared to other instruments currently in use (see table).
The Cedex Bio Bioprocess Analyzer is equipped with an automatic dilution function that extends the detection range and significantly reduces operator-induced deviations. On the Cedex Bio Bioprocess Analyzer, the sample does not need to be manually diluted before it is placed on the machine. Various photometric measurements (such as LDH [lactate dehydrogenase], IgG [immunoglobulin]) and ion-selective electrodes (sodium, potassium) are combined in the same instrument for flexible detection of individual samples.
Superior data quality of Cedex Bio Bioprocess Analyzer
Using the same reference standard (see blood glucose, lactic acid, and glutamine in Figure 1 ), the Cedex Bio bioprocess analyzer was compared to a mature instrument using enzyme membrane technology. Daytime parallel control experiments show that the Cedex Bio bioprocess analyzer has better accuracy and linearity. In addition, the high sensitivity of the Cedex Bio bioprocess analyzer makes it possible to have a nutrient-limited fermentation process. Moreover, subtle changes in metabolites during fermentation can be detected.
Results based on correlation studies in the field
Enzyme membrane analyzers require cumbersome maintenance, calibration, and frequent quality control. For the fermentation process of batch feeding of mammalian cells, the data quality of the Cedex Bio Bioprocess Analyzer correlation study is higher (see blood glucose and lactic acid in Figure 2 ), and it shows that the Cedex Bio bioprocess analyzer can be easily replaced. The enzyme membrane analyzer does not have any negative effects.
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Figure 2A: Correlation study results - blood glucose | Figure 2B: Correlation study results - lactic acid |
The Cedex Bio Bioprocess Analyzer analyzes sample product quality parameters such as LDH (representing release (ie, cytosolic protease)) and IgG (titer), which is a non-negligible advantage.
to sum up
The Cedex Bio Bioprocess Analyzer integrates the functionality of three devices on the same platform to perform multi-parameter testing of the same sample in minutes. Automated dilution reduces the number of steps that can be relied upon by the operator and reduces deviations. The easy-to-use and stable photometric, ion-selective and turbidity measurements are both reliable and reproducible, and can be directly compared between multiple instruments in different locations. The resulting sensitive, precise and accurate analytical data ensures a high level of control of the fermentation process.
English original text:
Using Roche Cedex Bio Analyzers for In-Process Monitoring in Biotech Production
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