Details
| Original language | English |
|---|---|
| Pages (from-to) | 55-63 |
| Number of pages | 9 |
| Journal | CYTOTECHNOLOGY |
| Volume | 6 |
| Issue number | 1 |
| Publication status | Published - May 1991 |
Abstract
An automated flow injection system for on-line analysis of proteins in real fermentation fluids was developed by combining the principles of stopped-flow, merging zones flow injection analysis (FIA) with antigen-antibody reactions. IgG in the sample reacted with its corresponding antibody (a-IgG) in the reagent solution. Formation of insoluble immunocomplexes resulted in an increase of the turbidity which was determined photometrically. This system was used to monitor monoclonal antibody production in high cell density perfusion culture of hybridoma cells. Perfusion was performed with a newly developed static filtration unit equipped with hydrophilic microporous tubular membranes. Different sampling devices were tested to obtain a cell-free sample stream for on-line product anlysis of high molecular weight (e.g., monoclonal antibodies) and low molecular weight (e.g., glucose, lactate) medium components. In fermentation fluids a good correlation (coefficient: 0.996) between the FIA method and an ELISA test was demonstrated. In a high density perfusion cultivation process mAb formation was succesfully monitored on-line over a period of 400 h using a reliable sampling system. Glucose and lactate were measured over the same period of time using a commercially available automatic analyser based on immobilized enzyme technology.
Keywords
- flow injection analysis, high cell density, hybridoma cells, on-line glucose and lactate measurements, on-line monitoring of monoclonal antibody formation, perfusion cultivation
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biotechnology
- Chemical Engineering(all)
- Bioengineering
- Engineering(all)
- Biomedical Engineering
- Biochemistry, Genetics and Molecular Biology(all)
- Clinical Biochemistry
- Biochemistry, Genetics and Molecular Biology(all)
- Cell Biology
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In: CYTOTECHNOLOGY, Vol. 6, No. 1, 05.1991, p. 55-63.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - On-line monitoring of monoclonal antibody formation in high density perfusion culture using FIA
AU - Fenge, Christel
AU - Fraune, Elisabeth
AU - Freitag, Ruth
AU - Scheper, Thomas
AU - Schügerl, Karl
PY - 1991/5
Y1 - 1991/5
N2 - An automated flow injection system for on-line analysis of proteins in real fermentation fluids was developed by combining the principles of stopped-flow, merging zones flow injection analysis (FIA) with antigen-antibody reactions. IgG in the sample reacted with its corresponding antibody (a-IgG) in the reagent solution. Formation of insoluble immunocomplexes resulted in an increase of the turbidity which was determined photometrically. This system was used to monitor monoclonal antibody production in high cell density perfusion culture of hybridoma cells. Perfusion was performed with a newly developed static filtration unit equipped with hydrophilic microporous tubular membranes. Different sampling devices were tested to obtain a cell-free sample stream for on-line product anlysis of high molecular weight (e.g., monoclonal antibodies) and low molecular weight (e.g., glucose, lactate) medium components. In fermentation fluids a good correlation (coefficient: 0.996) between the FIA method and an ELISA test was demonstrated. In a high density perfusion cultivation process mAb formation was succesfully monitored on-line over a period of 400 h using a reliable sampling system. Glucose and lactate were measured over the same period of time using a commercially available automatic analyser based on immobilized enzyme technology.
AB - An automated flow injection system for on-line analysis of proteins in real fermentation fluids was developed by combining the principles of stopped-flow, merging zones flow injection analysis (FIA) with antigen-antibody reactions. IgG in the sample reacted with its corresponding antibody (a-IgG) in the reagent solution. Formation of insoluble immunocomplexes resulted in an increase of the turbidity which was determined photometrically. This system was used to monitor monoclonal antibody production in high cell density perfusion culture of hybridoma cells. Perfusion was performed with a newly developed static filtration unit equipped with hydrophilic microporous tubular membranes. Different sampling devices were tested to obtain a cell-free sample stream for on-line product anlysis of high molecular weight (e.g., monoclonal antibodies) and low molecular weight (e.g., glucose, lactate) medium components. In fermentation fluids a good correlation (coefficient: 0.996) between the FIA method and an ELISA test was demonstrated. In a high density perfusion cultivation process mAb formation was succesfully monitored on-line over a period of 400 h using a reliable sampling system. Glucose and lactate were measured over the same period of time using a commercially available automatic analyser based on immobilized enzyme technology.
KW - flow injection analysis
KW - high cell density
KW - hybridoma cells
KW - on-line glucose and lactate measurements
KW - on-line monitoring of monoclonal antibody formation
KW - perfusion cultivation
UR - http://www.scopus.com/inward/record.url?scp=0026162116&partnerID=8YFLogxK
U2 - 10.1007/BF00353703
DO - 10.1007/BF00353703
M3 - Article
C2 - 1367401
AN - SCOPUS:0026162116
VL - 6
SP - 55
EP - 63
JO - CYTOTECHNOLOGY
JF - CYTOTECHNOLOGY
SN - 0920-9069
IS - 1
ER -