Bioprocess and Cell Culture

The C2MAP™ system measures component changes in a culture supernatant as culturing progresses using LC/MS/MS. This system can be used for a wide range of applications, from basic research of cell cultures including pluripotent stem cells (iPS cells and ES cells), mesenchymal stem cells, and antibody-producing cells, to scaling up of culture volumes and actual process development.

This Method Package enables the analysis of 125 compounds in under 20 minutes per sample. Performing analysis separately for each compound group such as amino acids and vitamins makes profiling of cell culture components very laborious, but with this method package a large number of culture medium components and secreted metabolites can be analyzed simultaneously. Compared to the previous version, several metabolites derived from amino acids, nucleic acids and the TCA cycle have been added, allowing even more comprehensive profiling information to be gathered.

A workflow that comprehensively evaluates commercially available T cell media using a Shimadzu LCMS-8050 triple quadrupole mass spectrometer is demonstrated in this application news. One hundred and twenty-five T cell culture medium components and secreted metabolites was analyzed simultaneously using a 17 minutes method. A list of compounds that differentiate the two commercially available T cell culture media were easily obtained using this method.

Optimization and control of cell culture processes are essential to increase production efficiency of biopharmaceuticals. In the field of cell therapy including regenerative medicines, enhanced control of the culture process is also becoming important to reduce cell variability and improve consistency of mass production of the cells. Comprehensive monitoring of culture supernatant tcomponents gives researchers useful information for these purposes.  However, current technologies for process monitoring are limited to measurement of pH, dissolving gases, and some small compounds such as glucose, glutamine, lactate, and ammonia in culture supernatant.

Application News No. C106A introduced an example of the monitoring of changes in culture supernatant components in accordance with the hybridoma cultivation process, using a high-performance liquid chromatograph (HPLC) coupled with a triple quadrupole (TQ) mass spectrometer (MS) and a cell culture profiling method package. This Application News introduces an example of the monitoring of changes in culture supernatant components in accordance with the iPS cellular cultivation process using a HPLC coupled with a quadrupole time-of-flight (Q-TOF) MS. Changes in the components in a culture supernatant can be monitored comprehensively using a combination of targeted SIM and non-targeted full scan analysis via a LC-Q-TOF-MS.

Development of technologies for the preparation and supply of iPS cells with high quality and a large amount is an essential requirement for commercialization of regenerative medicine. Conventionally, manual techniques such as Gene Expression analysis had been used to evaluate the status of cell differentiation, but because these are invasive techniques that cause cell disruption, they were generally applied as evaluation methods after culturing was completed. We developed the C2MAP cell culture media analysis platform using liquid chromatography-tandem mass spectrometry (LC-MS/MS) with the aim of establishing a non-invasive cell status evaluation method based on component analysis of the cell culture supernatant.

The growth condition and differentiation state (differentiated or undifferentiated) of cultured cells are important evaluation items in regenerative medicine and basic research. If these cell evaluations can be conducted in a simple manner by mass spectrometry, evaluation time and complicated work can be significantly reduced. Therefore, in order to examine the possibility of determining the state of cells by mass spectrometry, we conducted an automatic analysis of iPS cells and their culture media by AuraSolution™ using the Shimadzu MALDI-8020 benchtop linear MALDI-TOF mass spectrometer, and attempted a preliminary analysis by the statistical analysis software eMSTAT Solution™. The results are reported in this article.

This article introduces an example of cleaning validation of biopharmaceutical culture tanks using a Shimadzu on-line total organic carbon (TOC) analyzer. It is possible to minimize retention time and work between processes by using an on-line TOC analyzer. In particular, cleaning validation work can be substantially reduced by utilizing an on-line system because the culture tank rinse water is automatically sampled and measured periodically, and the acquired data are transferred successively to the upper level system. Use of this system in continuous production is also expected.

Until now, the ICP-MS and chelate determination techniques had been used in measurements of the metallic elements in the culture media. However, ICP-MS has high initial/running costs, and the complexity of the chelate determination method is a problem, as chelate determination requires sample pretreatment for each element in order to coordinate chelating agents (chelators) with high specificity for the target elements. Therefore, we measured the metallic elements in the culture media by atomic absorption spectrophotometry (AAS), which enables inexpensive and simple analysis of metallic elements. The content of this experiment is introduced in the following.

This article introduces an example of cleaning validation of biopharmaceutical culture tanks using a Shimadzu on-line total organic carbon (TOC) analyzer. It is possible to minimize retention time and work between processes by using an on-line TOC analyzer. In particular, cleaning validation work can be substantially reduced by utilizing an on-line system because the culture tank rinse water is automatically sampled and measured periodically, and the acquired data are transferred successively to the upper level system. Use of this system in continuous production is also expected.

This article introduces an example of cleaning validation of biopharmaceutical culture tanks using a Shimadzu on-line total organic carbon (TOC) analyzer. It is possible to minimize retention time and work between processes by using an on-line TOC analyzer. In particular, cleaning validation work can be substantially reduced by utilizing an on-line system because the culture tank rinse water is automatically sampled and measured periodically, and the acquired data are transferred successively to the upper level system. Use of this system in continuous production is also expected.

Developing an optimal cell culture bioprocess for the production of biopharmaceuticals requires routine monitoring of medium conditions such as pH, dissolved gas, carbon source (glucose) and nitrogen source (glutamine) for optimization and control of the cell culture process. However, culture medium is composed of various other biologically important compounds such as vitamins, amino acids, nucleic acids and other primary metabolites. Comprehensive analysis of these compounds would lead to more detailed understanding of the bioprocess. Recently, metabolomics has gained a lot of attention for cell culture profiling. In this study, we report comprehensive cell culture profiling of an iPS cell by metabolomics based LC-QTOFMS. SIM and Scan mode were simultaneously used in a single run for comprehensive metabolomics.