Document Detail


Amino acid analysis in mammalian cell culture media containing serum and high glucose concentrations by anion exchange chromatography and integrated pulsed amperometric detection.
MedLine Citation:
PMID:  15519579     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The direct separation detection of amino acids by anion exchange chromatography with integrated pulsed amperometric detection was optimized for the analysis of typical mammalian cell culture broth samples. Existing gradient elution conditions were adapted, considering the additions of peptone (2 g/L) and 10 vol% fetal calf serum to the medium as well as changing concentrations of glucose from 5.5 g/L up to complete consumption. Samples had to be analyzed in two dilutions with water (1:33.3 and 1:200) due to the strongly varying amino acid concentrations in the samples as a result of the medium composition and cell metabolism. The method was validated in a linear working range for the most common amino acids (2.5-7.5 and 1.25-3.75 microM for cystine/cysteine with 15 microl injection volume). The relative standard deviation of the method for all amino acids was less than 5%, with detection limits of less than 0.6 microM and quantitation limits of less than 1.6 microM. As an example, data for the amino acid composition of different media used for the production of inactivated influenza vaccines in cell culture are shown.
Authors:
Yvonne Genzel; Susanne König; Udo Reichl
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Analytical biochemistry     Volume:  335     ISSN:  0003-2697     ISO Abbreviation:  Anal. Biochem.     Publication Date:  2004 Dec 
Date Detail:
Created Date:  2004-11-02     Completed Date:  2005-03-17     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0370535     Medline TA:  Anal Biochem     Country:  United States    
Other Details:
Languages:  eng     Pagination:  119-25     Citation Subset:  IM    
Affiliation:
Max Planck Institute for Dynamics of Complex Technical Systems, Magdeburg, Sandtorstr. 1, 39106 Magdeburg, Germany. genzel@mpi-magdeburg.mpg.de
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MeSH Terms
Descriptor/Qualifier:
Amino Acids / analysis*
Animals
Cell Culture Techniques
Cells, Cultured / virology
Chromatography, High Pressure Liquid
Chromatography, Ion Exchange
Culture Media / chemistry*
Dogs
Glucose / metabolism*
Herpesvirus 4, Equid / chemistry*
Horses
Influenza A virus / chemistry
Sensitivity and Specificity
Serum / metabolism*
Chemical
Reg. No./Substance:
0/Amino Acids; 0/Culture Media; 50-99-7/Glucose

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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