ANALYSIS OF SUBSTANCES IN CELL CULTURE MEDIA WITH LCMSMS AND CELL CULTURE PROFILING METHOD PACKAGE

Study Objective:

Cells cultured in culture media are used in the biopharmaceutical industry during the development of active pharmaceutical ingredients such as monoclonal antibodies (MAbs) and vaccines, in basic research and subsequently in the production phase. The increase in cellular research in regenerative medicine and in the field of MAb-based drugs in the pharmaceutical market has highlighted the need to monitor culture media without damaging stem cells and MAb-producing cells. The fastest and easiest way to respond to this need is the simultaneous multi-component analysis of culture media components and metabolites secreted by cells using LC-MSMS. In this study, using the "Cell Culture Profiling Method Package" developed by Shimadzu (Kyoto, Japan), simultaneous analysis of 95 components (Table 1) was performed with an analysis time of 17 minutes per sample. Standard calibration curves were plotted for glucose and lactic acid for fully quantitative analysis, while for other components, semi-quantitative profile analysis was performed using 2-isopropylmalic acid internal standard (ISTD).

Table 1: Parameters that can be simultaneously monitored in 17-minute analysis time with Cell Culture Profiling Method Package

Methods and Materials: Cell Culture Conditions;

Standard preparation: Semi-Quantitative Analysis; 0.5 mmol/L 2-isopropylmalic acid solution (in Water:Acetonitrile (50:50, v/v)) is prepared.

Sample Preparation:

     

*Here, the MRM transitions of the parameters whose results are given in the application note are summarized. For MRM transitions of other parameters, please contact Ant Teknik Cihazlar Ltd.

Analytical Conditions: MS Conditions; Results and Evaluation:

In this study, glucose and lactic acid concentrations were determined quantitatively, while profile analysis (semi-quantitative analysis) was performed for the other 93 parameters, and the increase/decrease in the peak area of the target analyte relative to the area obtained from the internal standard used (2-isopropylmalic acid) was plotted and examined.

Chromatograms

Quantitative Analysis Results and Calibration Curves

Semi-Quantitative Analysis Results (Profile Analysis)

Information and Acknowledgements

The cell culture samples used in the study were provided by Marmara University Bioprocess Engineering and OMICs Research Group. We thank them for their support.   Dr. Engin Bayram Chemist/Deputy Head of MS Group Business Development & Application Ant Teknik Cihazlar    

References 1- Operating Guide; Shimadzu LC/MS/MS Method Package for Cell Culture Profiling, # 225-28413, February 2015. 2- Shimadzu ASMS Poster Presentation, Simultaneous analysis of primary metabolites by triple quadrupole LC/MS/MS using pentafluorophenylpropyl column, # PO-CON1447E, ASMS 2014. 3- Shimadzu Solutions for Biopharmaceuticals Analysis Application Databook, # C10G-E039A, March 2015.