Details
| Originalsprache | Englisch |
|---|---|
| Seiten (von - bis) | 130-138 |
| Seitenumfang | 9 |
| Fachzeitschrift | Engineering in life sciences |
| Jahrgang | 10 |
| Ausgabenummer | 2 |
| Publikationsstatus | Veröffentlicht - 12 Apr. 2010 |
Abstract
The improvement of specific productivity is a continuous challenge for bioprocesses involving mammalian cells, and hence, high-throughput methods and low-cost strategies are needed for the selection of high producers. The aim of this study was the productivity improvement of the hybridoma cell line IV F 19.23. For this purpose, a cell surface affinity matrix assay was established to identify and select high producers. This assay is based on the binding of secreted monoclonal antibodies to an affinity matrix assembled on the outer cell membrane. A protein microarray approach was used to investigate and optimize the functionality of the affinity matrix. The protein microarray was particularly useful to identify critical steps of the staining method, such as unspecific binding, before it was applied to the hybridoma cell line. Secreting hybridomas were treated with the affinity matrix and then selected via flow-cytometric cell sorting in four consecutive bulk sort rounds. The applied bulk strategy, allowing low screening costs, resulted in a 125% increase in specific productivity of the cell line in comparison to the initial population.
ASJC Scopus Sachgebiete
- Biochemie, Genetik und Molekularbiologie (insg.)
- Biotechnologie
- Umweltwissenschaften (insg.)
- Environmental engineering
- Chemische Verfahrenstechnik (insg.)
- Bioengineering
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in: Engineering in life sciences, Jahrgang 10, Nr. 2, 12.04.2010, S. 130-138.
Publikation: Beitrag in Fachzeitschrift › Artikel › Forschung › Peer-Review
}
TY - JOUR
T1 - Characterization and improvement of cell line performance via flow cytometry and cell sorting
AU - Moretti, Pierre
AU - Behr, Larissa
AU - Walter, Johanna G.
AU - Kasper, Cornelia
AU - Stahl, Frank
AU - Scheper, Thomas
PY - 2010/4/12
Y1 - 2010/4/12
N2 - The improvement of specific productivity is a continuous challenge for bioprocesses involving mammalian cells, and hence, high-throughput methods and low-cost strategies are needed for the selection of high producers. The aim of this study was the productivity improvement of the hybridoma cell line IV F 19.23. For this purpose, a cell surface affinity matrix assay was established to identify and select high producers. This assay is based on the binding of secreted monoclonal antibodies to an affinity matrix assembled on the outer cell membrane. A protein microarray approach was used to investigate and optimize the functionality of the affinity matrix. The protein microarray was particularly useful to identify critical steps of the staining method, such as unspecific binding, before it was applied to the hybridoma cell line. Secreting hybridomas were treated with the affinity matrix and then selected via flow-cytometric cell sorting in four consecutive bulk sort rounds. The applied bulk strategy, allowing low screening costs, resulted in a 125% increase in specific productivity of the cell line in comparison to the initial population.
AB - The improvement of specific productivity is a continuous challenge for bioprocesses involving mammalian cells, and hence, high-throughput methods and low-cost strategies are needed for the selection of high producers. The aim of this study was the productivity improvement of the hybridoma cell line IV F 19.23. For this purpose, a cell surface affinity matrix assay was established to identify and select high producers. This assay is based on the binding of secreted monoclonal antibodies to an affinity matrix assembled on the outer cell membrane. A protein microarray approach was used to investigate and optimize the functionality of the affinity matrix. The protein microarray was particularly useful to identify critical steps of the staining method, such as unspecific binding, before it was applied to the hybridoma cell line. Secreting hybridomas were treated with the affinity matrix and then selected via flow-cytometric cell sorting in four consecutive bulk sort rounds. The applied bulk strategy, allowing low screening costs, resulted in a 125% increase in specific productivity of the cell line in comparison to the initial population.
KW - Cell surface affinity matrix
KW - FACS
KW - Hybridoma
KW - Productivity
KW - Protein microarray
UR - http://www.scopus.com/inward/record.url?scp=77950837308&partnerID=8YFLogxK
U2 - 10.1002/elsc.200900076
DO - 10.1002/elsc.200900076
M3 - Article
AN - SCOPUS:77950837308
VL - 10
SP - 130
EP - 138
JO - Engineering in life sciences
JF - Engineering in life sciences
SN - 1618-0240
IS - 2
ER -