A new CHO (Chinese hamster ovary)-derived cell line expressing anti-TNFα monoclonal antibody with biosimilar potential

Abstract

Tumor necrosis factor alpha (TNFα) is a pro-inflammatory cytokine that mediates the homeostasis of immune responses; its exacerbated production is associated with the pathogenesis of autoimmune and chronic inflammatory diseases. Anti-TNFα drugs have revolutionized the treatment of inflammatory conditions such as rheumatoid arthritis and Crohn's disease. Currently, a worldwide race is on stage for the production of biosimilars moved by patent expiration of monoclonal antibodies (mAbs), such as anti-TNFα adalimumab. Our goal was to develop the first stage of an adalimumab biosimilar candidate with potential for national production, through the generation of a productive and stable cell line and assess its functionality. The robotic system ClonePix was used for screening and isolation of colonies from transfected CHO-S stable pools plated in semisolid medium. Selected clones were expanded based on growth and productivity. Purified mAbs from different clones were tested for binding and functional activity. The binding affinity of the denominated adabut clones to TNFα and FcRγ did not differ statistically when compared to reference adalimumab. One functional activity assay demonstrated the antibody neutralization capacity of the cytotoxicity induced by TNFα in L929 murine fibroblasts. A second assay confirmed adabut as an antagonist of the TNFα activity by the inhibition of the cell adhesion molecule expression in HUVEC cultures. The binding and functional activity analyses performed with selected adabut clones in comparison to reference adalimumab represent an important status of "non-inferiority," part of the process required for a biosimilar development. We generated and selected high-quality adabut clones which mAbs may be further developed as the first in-house made Brazilian biosimilar, demonstrating a success case for our incipient biotechnology industry, or also modified as biobetters, thus representing an innovative strategy for the patients' welfare.

Biobetters in patients with immune-mediated inflammatory disorders: An international Delphi consensus

“…Eight studies were excluded as they were not focused in the field of IMIDs. Finally, 7 studies were included in our systematic review [15][16][17][18][19][20][21]. Most studies were in vitro studies (6 [86%]) [15][16][17][18][19][20] followed by a phase I clinical trial (14%) [21] (Tables 1).…”

“…Finally, 7 studies were included in our systematic review [15][16][17][18][19][20][21]. Most studies were in vitro studies (6 [86%]) [15][16][17][18][19][20] followed by a phase I clinical trial (14%) [21] (Tables 1). All studies were of low quality according to the Newcastle Ottawa Scale (Table 2).…”

Functional production of human antibody by the filamentous fungus Aspergillus oryzae

“…Recently, the IgG production by CHO cells can generally reach 5 g/L through optimizing producer cell lines, culture media, and long incubation with high cell densities [37]. In addition, a screening of novel transfected CHO cells from 4345 clones has shown the adalimumab concentration on day 12 ranging from 124 to 594 mg/L by using the batch culture with glucose supplement [38]. In yeast species, the high-throughput screening was applied to select the P. pastoris strain producing IgGs up to 1 g/L in a 0.5 L bioreactor [39].…”

Functional production of human antibody by the filamentous fungus Aspergillus oryzae

“…Recently, the IgG production by CHO cells can generally reach 5 g/L through optimizing producer cell lines, culture media, and long incubation with high cell densities [37]. In addition, a screening of novel transfected CHO cells from 4345 clones has shown the adalimumab concentration on day 12 ranging from 124-594 mg/L by using the batch culture with glucose supplement [38]. In yeast species, the high-throughput screening was applied to select the P. pastoris strain producing IgGs up to 1 g/L in a 0.5 L bioreactor [39].…”