Purpose of the Study:
Cells grown in culture medium; They 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 increase in the area of MAb-based drugs in cellular research and in the pharmaceutical market in regenerative medicine fields has emerged as 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 to carry out these analyzes with LC-MSMS for the simultaneous multicomponent analysis of the culture medium components and metabolites secreted by the cells.
In this study, the simultaneous analysis of 95 components (Table-1) using ‘‘Cell Culture Profiling Method Package’’ developed by Shimadzu (Kyoto, Japan) was carried out for 17 minutes 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 internal standard (ISTD).
Table 1: Simultaneously with the Cell Culture Profiling Method Package in a 17-minute analysis period viewable parameters
Method and Materials:
Cell Culture Conditions;
Standard preparation:
Semi-Quantitative Analysis;
0.5 mmol / L of 2-isopropylmal acid solution (Water: Acetonitrile (50:50, v / v)) is prepared.
Sample Preparation:
* Here, the results of the application note, the parameters given, MRM transitions are summarized. You can contact us.
Analytical Conditions:
Conditions of MS;
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 the increase / decrease in the peak area of the target analyte was 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 (Bioprocess Engineering and OMICs Research Group) and 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 penta¬uorophenylpropyl column, # PO-CON1447E, ASMS 2014.
3- Shimadzu Solutions for Biopharmaceuticals Analysis Application Databook, # C10G-E039A, March 2015.
