LC-MS/MS AND ANALYSIS OF SUBSTANCES IN CELL CULTURE MEDIUM USING CELL CULTURE PROFILING METHOD PACKAGE

Purpose of the Study:

Cells grown in culture medium are used in the biopharmaceutical industry during the development of active pharmaceutical ingredients, such as monoclonal antibodies (MAbs) and vaccines, and in the production phase.

The expansion of MAb-based drugs in cellular research and in the pharmaceutical market in regenerative medicine fields has created the need to monitor culture media without damaging stem cells and MAb-producing cells.

The fastest and easiest way to address this need is to conduct these analyses using LC-MSMS for simultaneous multicomponent analysis of culture medium components and metabolites secreted by cells.

In this study, simultaneous analysis of 95 components (Table-1) was carried out using the "Cell Culture Profiling Method Package" developed by Shimadzu (Kyoto, Japan) with a 17-minute analysis time per sample.

For full quantitative analysis, standard calibration curves were drawn for glucose and lactic acid, and for other components, semi-quantitative profile analysis was performed using 2-isopropylmalic acid as the internal standard (ISTD).

Table 1: Parameters measurable simultaneously with the Cell Culture Profiling Method Package in a 17-minute analysis period

Method and Materials: Cell Culture Conditions;

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

Sample Preparation:

     

* Here, the application note results, the parameters provided, and MRM transitions are summarized. You can contact us.

Analytical Conditions: MS Conditions; Results and Evaluation

In this study, glucose and lactic acid concentrations were determined quantitatively, profile analysis (semi-quantitative analysis) was performed for the other 93 parameters, and increases/decreases in the peak area of the target analyte were investigated by plotting the area of the internal standard (2-isopropylmalic acid) used.

Chromatograms

Quantitative Analysis Results and Calibration Curves

Semi-Quantitative Analysis Results (Profile Analysis)

Information and Acknowledgments

Cell culture samples used in the study were provided by Marmara University Bioprocess Engineering and OMICs Research Group and we thank them for their support.

  Dr. Engin Bayram Chemist/MS Group Assistant Manager 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.