Efficient and reproducible generation of high-expressing, stable human cell lines without need for antibiotic selection
Open accessAbstract
BACKGROUND: Human cell lines are the most innovative choice of host cell for production of biopharmaceuticals since they allow for authentic posttranslational modification of therapeutic proteins. We present a new method for generating high and stable protein expressing cell lines based on human amniocytes without the requirement of antibiotic selection. RESULTS: Primary amniocytes from routine amniocentesis samples can be efficiently transformed with adenoviral functions resulting in stable human cell lines. Cotransfection of the primary human amniocytes with a plasmid expressing adenoviral E1 functions plus a second plasmid containing a gene of interest resulted in permanent cell lines expressing up to 30 pg/cell/day of a fully glycosylated and sialylated protein. Expression of the gene of interest is very stable for more than 90 passages and, importantly, was achieved in the absence of any antibiotic selection. CONCLUSION: We describe an improved method for developing high protein expressing stable human cell lines. These cell lines are of non-tumor origin, they are immortalized by a function not oncogenic in human and they are from an ethically accepted and easily accessible cell source. Since the cell can be easily adapted to growth in serum-free and chemically defined medium they fulfill the requirements of biopharmaceutical production processes.
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CAP and HuH-7 cells display human-like glycosylation profiles.
“Two other cell lines, the CAP (CEVEC's Amniocyte Production) cells of human amniocytes origin and the HuH-7 cells of human hepatocellular carcinoma origin, are presently tested for recombinant glycoprotein production in preclinical phases and both display human-like glycosylation profiles (Schiedner et al., 2008;Wolfel et al., 2011;Enjolras et al., 2012;Swiech et al., 2012;Wissing et al., 2015).”
From Therapeutic glycoprotein production in mammalian cells (2017) · cites this paper as Schiedner et al., 2008
The Results section (page unavailable) reports that hAAT produced in human amniocyte cell lines was glycosylated and sialylated, with test results comparable to hAAT from human serum. This supports a human-like feature for the amniocyte-derived cells, but the paper does not establish a full glycosylation profile for CAP cells or address HuH-7 cells. (LLM, conf=0.86)
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Therapeutic glycoprotein production in mammalian cells
Lalonde, M.-E., & Durocher, Y. (2017) · Journal of Biotechnology · doi:10.1016/j.jbiotec.2017.04.028 · Cited by 296 · see citing works
- Authors
- Schiedner, G., Hertel, S., Bialek, C., Kewes, H., Waschütza, G., & Volpers, C.
- Affiliations
- Federal Office of Administration (DE)
- Corresponding author
- Schiedner, Gudrun
- Venue
- BMC Biotechnology
- Published
- 2008
- Volume
- 8
- Issue
- 1
- Pages
- 13
- ISSN
- 1472-6750 (ISSN-L)
- PMC
- PMC2262890
- OpenAlex
- W2137488699
- Cited by
- 50
- References
- 0
- Citations by year
- 2026: 1 · 2025: 2 · 2024: 1 · 2023: 1 · 2022: 1 · 2021: 1 · 2020: 2 · 2019: 8 · 2018: 2 · 2017: 5 · 2016: 2 · 2015: 5 · 2014: 5 · 2013: 5 · 2012: 5
- FWCI
- 0.55
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- cc-by
- Locations
- BMC Biotechnology (publishedVersion, cc-by)
- Europe PMC (PubMed Central) (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Funders
- Bundesministerium für Bildung und Forschung
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Virus-based gene therapy research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein purification and stability
- Keywords
- human cell lines, amniocytes, cotransfection, post-translational modification, biopharmaceutical manufacturing, cell immortalization
- MeSH terms
- Amniotic Fluid, Anti-Bacterial Agents, Cell Line, Humans, Recombinant Proteins, Transfection, Protein Engineering, Cell Culture Techniques, Genetic Enhancement, Cell Proliferation
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