Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
Review
. 2022 Mar 4:10:856049.
doi: 10.3389/fbioe.2022.856049. eCollection 2022.

Strategies and Considerations for Improving Recombinant Antibody Production and Quality in Chinese Hamster Ovary Cells

Jun-He Zhang  1   2   3 , Lin-Lin Shan  2 , Fan Liang  1 , Chen-Yang Du  1 , Jing-Jing Li  2
Affiliations
Review

Strategies and Considerations for Improving Recombinant Antibody Production and Quality in Chinese Hamster Ovary Cells

Jun-He Zhang et al. Front Bioeng Biotechnol. .

Abstract

Recombinant antibodies are rapidly developing therapeutic agents; approximately 40 novel antibody molecules enter clinical trials each year, most of which are produced from Chinese hamster ovary (CHO) cells. However, one of the major bottlenecks restricting the development of antibody drugs is how to perform high-level expression and production of recombinant antibodies. The high-efficiency expression and quality of recombinant antibodies in CHO cells is determined by multiple factors. This review provides a comprehensive overview of several state-of-the-art approaches, such as optimization of gene sequence of antibody, construction and optimization of high-efficiency expression vector, using antibody expression system, transformation of host cell lines, and glycosylation modification. Finally, the authors discuss the potential of large-scale production of recombinant antibodies and development of culture processes for biopharmaceutical manufacturing in the future.

Keywords: Chinese hamster ovary cells; expression vector; genetic engineering; glycosylation; recombinant antibody.

PubMed Disclaimer

Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Development process of recombinant antibody drugs. Mouse monoclonal antibodies are all mouse-derived components, with large side effects and high immunogenicity; human-mouse chimeric antibodies are 65% human-derived components, with lower side effects than mouse monoclonal antibody; human-derived components in humanized antibodies are over 90%, with mild side effects and very low immunogenicity; fully human antibodies are all human-derived components, which can obviously remove the immunogenicity and side effects, and with good therapeutic effect.
FIGURE 2
FIGURE 2
A workflow for optimizing recombinant antibody production and quality in CHO cells. Too many factors can impact recombinant antibody production and quality in CHO cells, therefore, it is important to follow a specific workflow when dealing with recombinant antibody production and quality.
FIGURE 3
FIGURE 3
Basic structure of antibody. Each IgG antibody molecule consists of four polypeptide chains (two identical light chains and two identical heavy chains joined by disulfide bonds) and has two antigen-binding sites. Each light chain and each heavy chain consists of a variable region and a constant region. Each heavy chain consists of a variable domain (VH) and three or four constant domains (CH); each light chain consists of a variable domain (VL) and a single constant domain (CL). Human IgG structure with glycans attached at Asn297 N-glycosylation site in the CH2.
FIGURE 4
FIGURE 4
Types of glycosylation modification of recombinant antibodies. The representative N-glycan structures identified on antibody Fc fragment are G0F, G1F, and G2F. G0F: asialo, agalactose, biantennary complex, core substituted with fucose; G1F: asialo, mono-galactosylated, biantennary complex, core substituted with fucose; G2F: asialo, galactosylated, biantennary complex, core substituted with fucose.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Ahmadi M., Mahboudi F., Akbari Eidgahi M. R., Nematpour F., Ahmadi S., Ebadat S., et al. (2016). Evaluating the Efficiency of phiC31 Integrase-Mediated Monoclonal Antibody Expression in CHO Cells. Biotechnol. Prog. 32, 1570–1576. 10.1002/btpr.2362 - DOI - PubMed
    1. Akbarzadeh-Sharbaf S., Yakhchali B., Minuchehr Z., Shokrgozar M. A., Zeinali S. (2013). Expression Enhancement in Trastuzumab Therapeutic Monoclonal Antibody Production Using Genomic Amplification with Methotrexate. Avicenna J. Med. Biotechnol. 5, 87–95. - PMC - PubMed
    1. Alessandri L., Ouellette D., Acquah A., Rieser M., Leblond D., Saltarelli M., et al. (2012). Increased Serum Clearance of Oligomannose Species Present on a Human IgG1 Molecule. MAbs 4, 509–520. 10.4161/mabs.20450 - DOI - PMC - PubMed
    1. An Z. Q. (2009). Therapeutic Monoclonal Antibodies: From Bench to Clinic. New Jersey: John Wiley & Sons, 73–75.
    1. Attallah C., Aguilar M. F., Forno G., Etcheverrigaray M., Brigido M. D. M., Maranhão A. Q., et al. (2020). The Glycosylation of Anti-rhIFN-α2b Recombinant Antibodies Influences the Antigen-Neutralizing Activity. Biotechnol. Lett. 42, 1369–1381. 10.1007/s10529-020-02879-0 - DOI - PubMed

LinkOut - more resources