EN
Developing selection strategy for CHO-K1 cell line that secretes scfv-Fc fusion antibodies using ClonePix2
Öz
In the pharmaceutical industry, biopharmaceuticals (biologics) are gaining market share. There has been a dramatic increase in the sale and market penetration of monoclonal antibodies in particular. Typically, therapeutic antibodies are produced using high-expression, clonal, or recombinant CHO cell lines. CHO cells dominate the market as a commercial production host due to their ease of use, built-in regulatory records, and security profiles. While traditional limiting-dilution and cloning-ring regulations are frequently used to select mammalian cell lines that produce high levels of proteins, they have a number of drawbacks. ClonePix2 is a fully automated, single cell-based clone selector that significantly increases the likelihood of rapidly selecting high-production clones with high monoclonality. Scfv-Fc recombinant antibody structures with a variety of therapeutic advantages have gained prominence in recent years. Single cell cloning of CHO cells expressing the scfv-Fc fusion protein, which differs from the classical immunoglobulin structure, was performed in situ using the ClonePix2 device using FITC-tagged anti-Fc and anti-H+L antibodies. The fluorescent intensity parameters of the resulting cell clones were analyzed. Additionally, ELISA was used to determine the production capacities of the best clones. As a result, it was established that anti-Fc antibody recognizes the scfv-Fc fusion protein in a semi-solid environment, enabling the identification of higher production clones.
Anahtar Kelimeler
Destekleyen Kurum
This work has been supported by the TÜBİTAK, Marmara Research Center, Medical Biotechnology Department of Life Sciences.
Teşekkür
I would like to express my gratitude to Esin Bayralı, Gökhan Güler and Evrim Tekel for their excellent technical assistanc, as well as to Koray B. Balcioglu, H. Ümit Öztürk, Hivda Ülbeği Polat, Müge Serhatlı, Hilal Yazıcı Malkoçoğlu and Melis Denizci Öncü for their comments on this article.
Kaynakça
- Wurm, F. M., Production of recombinant protein therapeutics in cultivated mammalian cells. Nature Biotechnology, 2004. 22(11): p. 1393–1398.
- Dharshanan, S. and C. S. Hung, Screening and subcloning of high producer transfectomas using semisolid media and automated colony picker. Methods Mol. Biology, 2014. 1131: p. 105–112.
- Nakamura, T. and T. Omasa, Optimization of cell line development in the GS-CHO expression system using a high-throughput, single cell-based clone selection system. Jornal of Bioscience and Bioengineering, 2015. 120(3): p. 323–329.
- Tsuruta L.R., et al., Genetic analyses of Per.C6 cell clones producing a therapeutic monoclonal antibody regarding productivity and long-term stability. Applied Genetics and Molecular Biotechnology, 2016. 100: p. 10031–11041.
- Tejwani, V., et al., High-throughput and automation advances for accelerating single-cell cloning, monoclonality and early phase clone screening steps in mammalian cell line development for biologics production. Biotechnol. Prog., 2021. 37(6): p. 3208.
- O’Rourke, S. M., et al., Robotic selection for the rapid development of stable CHO cell lines for HIV vaccine production. PLoS One, 2018. 13 (8): p.1–22.
- Ahmad, Z. A., et al., ScFv antibody: Principles and clinical application. Journal of Immunology Research, 2012. 2012: p. 1-15.
- Yu, J., Y. Song, and W. Tian, How to select IgG subclasses in developing anti-tumor therapeutic antibodies. Journal of Hematology and Oncology, 2020. 13(45): p.1-10.
Ayrıntılar
Birincil Dil
İngilizce
Konular
Biyokimya ve Hücre Biyolojisi (Diğer), Endüstriyel Biyoteknoloji
Bölüm
Araştırma Makalesi
Yazarlar
Yayımlanma Tarihi
15 Aralık 2022
Gönderilme Tarihi
6 Mayıs 2022
Kabul Tarihi
2 Eylül 2022
Yayımlandığı Sayı
Yıl 2022 Cilt: 5 Sayı: 3
APA
Özdemir Bahadır, A. (2022). Developing selection strategy for CHO-K1 cell line that secretes scfv-Fc fusion antibodies using ClonePix2. International Journal of Life Sciences and Biotechnology, 5(3), 533-545. https://doi.org/10.38001/ijlsb.1112823
AMA
1.Özdemir Bahadır A. Developing selection strategy for CHO-K1 cell line that secretes scfv-Fc fusion antibodies using ClonePix2. Int J. Life Sci. Biotechnol. 2022;5(3):533-545. doi:10.38001/ijlsb.1112823
Chicago
Özdemir Bahadır, Aylin. 2022. “Developing selection strategy for CHO-K1 cell line that secretes scfv-Fc fusion antibodies using ClonePix2”. International Journal of Life Sciences and Biotechnology 5 (3): 533-45. https://doi.org/10.38001/ijlsb.1112823.
EndNote
Özdemir Bahadır A (01 Aralık 2022) Developing selection strategy for CHO-K1 cell line that secretes scfv-Fc fusion antibodies using ClonePix2. International Journal of Life Sciences and Biotechnology 5 3 533–545.
IEEE
[1]A. Özdemir Bahadır, “Developing selection strategy for CHO-K1 cell line that secretes scfv-Fc fusion antibodies using ClonePix2”, Int J. Life Sci. Biotechnol., c. 5, sy 3, ss. 533–545, Ara. 2022, doi: 10.38001/ijlsb.1112823.
ISNAD
Özdemir Bahadır, Aylin. “Developing selection strategy for CHO-K1 cell line that secretes scfv-Fc fusion antibodies using ClonePix2”. International Journal of Life Sciences and Biotechnology 5/3 (01 Aralık 2022): 533-545. https://doi.org/10.38001/ijlsb.1112823.
JAMA
1.Özdemir Bahadır A. Developing selection strategy for CHO-K1 cell line that secretes scfv-Fc fusion antibodies using ClonePix2. Int J. Life Sci. Biotechnol. 2022;5:533–545.
MLA
Özdemir Bahadır, Aylin. “Developing selection strategy for CHO-K1 cell line that secretes scfv-Fc fusion antibodies using ClonePix2”. International Journal of Life Sciences and Biotechnology, c. 5, sy 3, Aralık 2022, ss. 533-45, doi:10.38001/ijlsb.1112823.
Vancouver
1.Aylin Özdemir Bahadır. Developing selection strategy for CHO-K1 cell line that secretes scfv-Fc fusion antibodies using ClonePix2. Int J. Life Sci. Biotechnol. 01 Aralık 2022;5(3):533-45. doi:10.38001/ijlsb.1112823