Details
| Original language | English |
|---|---|
| Pages (from-to) | 288-293 |
| Number of pages | 6 |
| Journal | Engineering in life sciences |
| Volume | 21 |
| Issue number | 5 |
| Early online date | 6 Jan 2021 |
| Publication status | Published - May 2021 |
Abstract
Due to the increasing economic and social relevance of biotherapeutics, their production processes are continually being reconsidered and reoptimized in an effort to secure higher product concentrations and qualities. Monitoring the productivity of cultured cells is therefore a critically important part of the cultivation process. Traditionally, this is achieved by determining the overall product titer by high performance liquid chromatography (HPLC), and then calculating the specific cell productivity based on this titer and an associated viable cell density. Unfortunately, this process is typically time-consuming and laborious. In this study, the productivity of Chinese Hamster Ovary (CHO) cells expressing a monoclonal antibody was analyzed over the course of the cultivation process. In addition to calculating the specific cell productivity based on the traditional product titer determined by HPLC analysis, culture productivity of single cells was also analyzed via flow cytometry using a cold capture assay. The cold capture assay is a cell surface labelling technique described by Brezinsky et al., which allows for the visualization of a product on the surface of the producing cell. The cell productivity results obtained via HPLC and the results of cold capture assay remained in great accordance over the whole cultivation process. Accordingly, our study demonstrates that the cold capture assay offers an interesting, comparatively time-effective, and potentially cheaper alternative for monitoring the productivity of a cell culture.
Keywords
- CHO cells, mammalian cell culture, monoclonal antibody production, secretion assay, specific cell productivity
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biotechnology
- Environmental Science(all)
- Environmental Engineering
- Chemical Engineering(all)
- Bioengineering
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In: Engineering in life sciences, Vol. 21, No. 5, 05.2021, p. 288-293.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Monitoring cell productivity for the production of recombinant proteins by flow cytometry
T2 - An effective application using the cold capture assay
AU - Meyer, Katharina V.
AU - Siller, Ina G.
AU - Schellenberg, Jana
AU - Gonzalez Salcedo, Alina
AU - Solle, Dörte
AU - Matuszczyk, Jens
AU - Scheper, Thomas
AU - Bahnemann, Janina
N1 - Funding Information: The authors acknowledge the financial support of the German Research Foundation (DFG) via the Emmy Noether Programme (346772917). Furthermore the authors would like to thank the Open Access fund of Leibniz Universität Hannover for the funding of the publication of this article.
PY - 2021/5
Y1 - 2021/5
N2 - Due to the increasing economic and social relevance of biotherapeutics, their production processes are continually being reconsidered and reoptimized in an effort to secure higher product concentrations and qualities. Monitoring the productivity of cultured cells is therefore a critically important part of the cultivation process. Traditionally, this is achieved by determining the overall product titer by high performance liquid chromatography (HPLC), and then calculating the specific cell productivity based on this titer and an associated viable cell density. Unfortunately, this process is typically time-consuming and laborious. In this study, the productivity of Chinese Hamster Ovary (CHO) cells expressing a monoclonal antibody was analyzed over the course of the cultivation process. In addition to calculating the specific cell productivity based on the traditional product titer determined by HPLC analysis, culture productivity of single cells was also analyzed via flow cytometry using a cold capture assay. The cold capture assay is a cell surface labelling technique described by Brezinsky et al., which allows for the visualization of a product on the surface of the producing cell. The cell productivity results obtained via HPLC and the results of cold capture assay remained in great accordance over the whole cultivation process. Accordingly, our study demonstrates that the cold capture assay offers an interesting, comparatively time-effective, and potentially cheaper alternative for monitoring the productivity of a cell culture.
AB - Due to the increasing economic and social relevance of biotherapeutics, their production processes are continually being reconsidered and reoptimized in an effort to secure higher product concentrations and qualities. Monitoring the productivity of cultured cells is therefore a critically important part of the cultivation process. Traditionally, this is achieved by determining the overall product titer by high performance liquid chromatography (HPLC), and then calculating the specific cell productivity based on this titer and an associated viable cell density. Unfortunately, this process is typically time-consuming and laborious. In this study, the productivity of Chinese Hamster Ovary (CHO) cells expressing a monoclonal antibody was analyzed over the course of the cultivation process. In addition to calculating the specific cell productivity based on the traditional product titer determined by HPLC analysis, culture productivity of single cells was also analyzed via flow cytometry using a cold capture assay. The cold capture assay is a cell surface labelling technique described by Brezinsky et al., which allows for the visualization of a product on the surface of the producing cell. The cell productivity results obtained via HPLC and the results of cold capture assay remained in great accordance over the whole cultivation process. Accordingly, our study demonstrates that the cold capture assay offers an interesting, comparatively time-effective, and potentially cheaper alternative for monitoring the productivity of a cell culture.
KW - CHO cells
KW - mammalian cell culture
KW - monoclonal antibody production
KW - secretion assay
KW - specific cell productivity
UR - http://www.scopus.com/inward/record.url?scp=85099059443&partnerID=8YFLogxK
U2 - 10.1002/elsc.202000049
DO - 10.1002/elsc.202000049
M3 - Article
AN - SCOPUS:85099059443
VL - 21
SP - 288
EP - 293
JO - Engineering in life sciences
JF - Engineering in life sciences
SN - 1618-0240
IS - 5
ER -