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Human cell lines: A promising alternative for recombinant FIX production
Aline de Sousa Bomfim1, Marcela Cristina Corrêa de Freitas2, Virgínia Picanço-Castro3
1Department of Clinical, Toxicological and Food Science Analysis, Faculty of Pharmaceutical Sciences of Ribeirão Preto, University of São Paulo, 2501 Tenente Catão Roxo Street 14051-140, Ribeirão Preto, SP, Brazil; Center for Cell-based Therapy and Regional Blood Center, University of São Paulo, 2501 Tenente Catão Roxo Street 14051-140, Ribeirão Preto, SP, Brazil.
SK-Hep-1 cells are effective for producing active recombinant human Factor IX (rhFIX), a key treatment for Hemophilia B. This study highlights SK-Hep-1 as a promising platform for rhFIX biopharmaceutical production.
Area of Science:
- Biotechnology
- Biochemistry
- Cell Biology
Background:
- Factor IX (FIX) is a crucial vitamin K-dependent protein used in Hemophilia B treatment.
- Recombinant human FIX (rhFIX) production requires efficient expression and biological activity.
Purpose of the Study:
- To evaluate recombinant human FIX (rhFIX) expression in 293T and SK-Hep-1 human cell lines.
- To identify optimal cell lines and conditions for producing biologically active rhFIX.
Main Methods:
- Lentiviral integration analysis in 293T-FIX cells.
- Gene expression analysis of γ-carboxylation pathway genes (VKORC1, carboxylase).
- rhFIX expression and activity assessment in different cell lines and media conditions (Ca+2, Mg+2 enrichment).
Main Results:
- SK-Hep-1-FIX cells produced higher levels of biologically active rhFIX compared to 293T-FIX cells.
- Despite lower VKORC1 and carboxylase expression, SK-Hep-1 cells secreted significantly more active rhFIX.
- 293T cells showed efficient carboxylation potential but lower overall active rhFIX yield (28% active).
- Ca+2 and Mg+2 enrichment enhanced rhFIX concentration in 293T cells but not in SK-Hep-1 cells.
Conclusions:
- The SK-Hep-1 cell line is a highly effective platform for producing biologically active rhFIX.
- SK-Hep-1 offers a novel biotechnological platform for recombinant protein production, particularly for therapeutic proteins like rhFIX.
