Shimadzu Europa
High-Throughput Multicomponent Profiling of Cell Culture Medium by LC/MS/MS for Advance Process Monitoring of Biopharmaceutical Products

Skip to Navigation

Advertorial

  • Published: Nov 1, 2017
  • Source: Shimadzu Europa GmbH
  • Categories: Base Peak
thumbnail image: <font size=3>Shimadzu Europa</font><br />High-Throughput Multicomponent Profiling of Cell Culture Medium by LC/MS/MS for Advance Process Monitoring of Biopharmaceutical Products

Authors: 1Gurmil Gendeh, 1Tairo Ogura, 2Takashi Suzuki, 2Masaroshi Takahashi
1Shimadzu Scientific Instruments, Columbia, MD and 2Shimadzu Corporation, Kyoto, Japan
www.shimadzu.eu, shimadzu@shimadzu.eu

Introduction

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 media also consist of various other biologically important compounds such as vitamins, nucleic acids and other primary metabolites, which would lead to more detailed understanding of the bioprocess if monitored altogether.

To meet the demand for comprehensive analysis of medium component, we optimized the analytical conditions and developed a high-throughput LC-MS/MS “Method Package for Cell Culture Profiling” that can monitor relative abundance of 95 compounds listed herein at a rate of 17 minutes per sample. Using this Method Package, we demonstrated the change in abundance of culture medium components associated with hybridoma growth over a period of 5 days.

To our best knowledge, the method represents the highest number of cell culture medium components and their secreted metabolites analyzed simultaneously in a single analysis.

.

.

Materials and Methods

A murine hybridoma cell line was cultured in DMEM (see Table 1 for conditions) and its culture supernatant was sampled every 24 hours for 5 days after inoculation. LCMS sample was prepared by adding an internal standard to the sample and then removing proteins by taking supernatant after mixing with acetonitrile, which was further diluted with ultrapure water prior to injection. 1 µL was injected to LCMS for simultaneous MRM quantitation of all 96 compounds. Fig. 1 shows a growth curve and viability plot of the cell line, and Fig. 2 shows the quantitative value (ratio of peak area with respect to internal standard) of representative compounds over 5 days.

Instrument: As an LC-MS/MS system, UHPLC was coupled to triple quadrupole mass spectrometer (Nexera MP with LCMS-8050, Shimadzu Corporation, Kyoto, Japan). LC-MS/MS with electrospray ionization was operated in multiple-reaction-monitoring (MRM) mode with ultra-fast polarity switching.

.

Shimadzu LCMS-8050 High Speed Triple Quadrupole Mass Spectrometer with
Ultra Fast Polarity Switching (5 msec) and Ultra Fast MRM (Max. 555 transition/sec)

.

.

.

Results

Representative results are shown below. (A) Glucose, glutamine and few other amino acids, which are the primary sources of carbon and nitrogen, have decreased in abundance with growing cell number. (B) In contrast, lactic acid increased in abundance over time as a result of glucose consumption for anaerobic respiration. Similar pattern of increase was observed for a few other compounds. (C) No change in relative abundance was observed for essential amino acids and some vitamins.

.

.

Conclusion

“Method Package for Cell Culture Profiling” enables users to analyze simultaneously multicomponents of cell culture medium using UHPLC coupled with triple quadrupole mass spectrometry. This package covers analysis method not only for basal medium of cell culture but also metabolites secreted by cells. And the optimized method realizes simultaneous analysis in 17 minutes that include chromatographic time and column equilibration time. This package supports simultaneous analysis of high concentration components and trace components in single analysis.

For more information please click here

For Research Use Only. Not for use in diagnostic procedure.

This publication may contain references to products that are not available in your country. Please contact us to check the availability of these products in your country.

The content of this publication shall not be reproduced, altered or sold for any commercial purpose without the written approval of Shimadzu. Company names, product/service names and logos used in this publication are trademarks and trade names of Shimadzu Corporation or its affiliates, whether or not they are used with trademark symbol “TM” or “®”. Third-party trademarks and trade names may be used in this publication to refer to either the entities or their products/services. Shimadzu disclaims any proprietary interest in trademarks and trade names other than its own. The information contained herein is provided to you "as is" without warranty of any kind including without limitation warranties as to its accuracy or completeness. Shimadzu does not assume any responsibility or liability for any damage, whether direct or indirect, relating to the use of this publication. This publication is based upon the information available to Shimadzu on or before the date of publication, and subject to change without notice.

Social Links

Share This Links

Bookmark and Share

Microsites

Suppliers Selection
Societies Selection

Banner Ad

Banner Ad

Most Viewed

Copyright Information

Interested in separation science? Visit our sister site separationsNOW.com

Copyright © 2017 John Wiley & Sons, Inc. All Rights Reserved