Page 14 - Pharmaceutical- Guide to Biopharmaceutical

Culture

Monitoring Components in Cell Culture Solutions Nexera X3 UHPLC + LCMS-8060 + Cell Culture Profiling

Simultaneous Analysis of Components in CHO Cell Culture benefits

Supernatant for Optimization of the Culture Process Cell Line Optimization
click here • Simultaneous analysis of up to 125 amino acids, vitamins, nucleic acids, or other
compounds contained in the culture supernatant within 20 minutes.
Operating Principle and Features Measurement Method and Results
• Sensitivity levels have been specified based on the concentration of target components
Triple Quad LCMS-8060 systems feature UF technology that enables CHO cell culture solutions were sampled every 24 hours and being measured, which can reduce the work involved in creating a series of dilutions.
both high sensitivity and high speed. Due to high-speed scanning and supernatants obtained by centrifugation. The supernatant was
high-speed positive-negative ionization switching capability, the system deproteinized and diluted with ultrapure water before analysis. • Time-series monitoring of multiple components provides powerful support for Culture
can simultaneously analyze multiple metabolites with a wide variety Analytical results over a time course are graphically represented in Fig. 2. optimizing cultivation parameters.
of chemical properties. The cell culture profiling method package is an Important cell culture factors such as consumption of nutrients during
analysis method optimized for analyzing multiple components in culture cell proliferation and metabolic secretion can be monitored.
supernatant solutions. It enables analyzing up to 125 compounds (refer
to product specifications) in 20 minutes or less (Fig. 1).
This example describes monitoring the components in CHO cell culture
supernatant over time.
(x1,000,000)
2.00 Purification

1.50 Argininosuccinic acid Phenylalanine
Hexose (Glucose) 5-Oxoproline Putrescine Valine Tyrosine Isoleucine
1.00 Aspartic acid Orotic acid Histidine Arginine 1-Methylhistidine Choline Pantothenic acid Tryptophan

0.50 Threonine Uridine Methionine Cytidine Leucine Characterization

0.00
1.0 2.0 3.0 4.0 5.0 6.0 7.0 8.0 min
Fig. 1 MRM Chromatogram
Hexose (Glucose) Lactic acid Folic acid
Area ratio Area ratio Area ratio Specifications LC/MS/MS Method Package for Cell Culture Profiling Ver.2 Quality Control
Software
LC unit Nexera X3 (SCL-40, LC-40BX3, CTO-40S, SIL-40C X3, MR20 µL mixer)
day day day
Asparagine Pyridoxal Arginine MS unit LCMS-8045/-8050/-8060
Analysis cycle Less than 20 minutes per sample
Area ratio Area ratio Area ratio Registered compounds 125 compounds + internal standard substance (2-isopropylmalic acid)
Amino acids and metabolites
Nucleic acids and metabolites 60 compounds
31 compounds
day day day Vitamins 15 compounds
Sugars 4 compounds Pharmacokinetics
Fig. 2 Time-Series Changes in Each Component Other (organic acids, etc.) 15 compounds
Separation mode Reversed
Conclusion Application Examples
Detection mode MRM (positive/negative)
Rapid and comprehensive cell culture media analysis for the • Spent media analysis
determination of nutrients and metabolites is possible. • Qualitative and quantitative analysis of culture media, bovine serum,
and other samples (requires calibration curve preparation)
• Metabolomic analysis of culture supernatant and body fluids Others

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