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ID: 207797.0, MPI für Dynamik komplexer technischer Systeme / Bioprocess Engineering
Amino acid analysis in mammalian cell culture media containing serum and high glucose concentrations by anion exchange chromatography and integrated pulsed amperometric detection
Authors:Genzel, Y.; König, S.; Reichl, U.
Date of Publication (YYYY-MM-DD):2004
Title of Journal:Analytical Biochemistry
Start Page:119
End Page:125
Review Status:not specified
Audience:Experts Only
Intended Educational Use:No
Abstract / Description: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 μM for cystine/cysteine with 15 μl 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 μM and quantitation limits of less than 1.6 μM. 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.
External Publication Status:published
Document Type:Article
Communicated by:Udo Reichl
Affiliations:MPI für Dynamik komplexer technischer Systeme/Bioprocess Engineering
Identifiers:LOCALID:1007 24/04
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