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Evaluation of changes in promoters, use of UCOES and chain order to improve the antibody production in CHO cells.
Rocha-Pizaña, Maria Del Refugio; Ascencio-Favela, Guadalupe; Soto-García, Brenda Maribell; Martinez-Fierro, Margarita de la Luz; Alvarez, Mario Moisés.
Affiliation
  • Rocha-Pizaña MDR; Centro de Biotecnología-FEMSA, Tecnológico de Monterrey at Monterrey, Ave. Eugenio Garza Sada 2501 Sur Col. Tecnológico, CP 64849, Monterrey, Nuevo León, Mexico.
  • Ascencio-Favela G; Centro de Biotecnología-FEMSA, Tecnológico de Monterrey at Monterrey, Ave. Eugenio Garza Sada 2501 Sur Col. Tecnológico, CP 64849, Monterrey, Nuevo León, Mexico.
  • Soto-García BM; Centro de Biotecnología-FEMSA, Tecnológico de Monterrey at Monterrey, Ave. Eugenio Garza Sada 2501 Sur Col. Tecnológico, CP 64849, Monterrey, Nuevo León, Mexico.
  • Martinez-Fierro ML; Molecular Medicine Laboratory, Unidad Academica de Medicina Humana y Ciencias de la Salud de la Universidad Autonoma de Zacatecas, Zacatecas, Mexico.
  • Alvarez MM; Centro de Biotecnología-FEMSA, Tecnológico de Monterrey at Monterrey, Ave. Eugenio Garza Sada 2501 Sur Col. Tecnológico, CP 64849, Monterrey, Nuevo León, Mexico. Electronic address: mario.alvarez@itesm.mx.
Protein Expr Purif ; 132: 108-115, 2017 04.
Article in En | MEDLINE | ID: mdl-28161546
Therapy with biopharmaceuticals, mainly recombinant antibodies, offers patients higher life expectancy and better life quality than pharmacologic therapy. Countries with the highest scientific development are investing in this kind of therapy, and this is why the optimization of the production of these recombinant proteins would lead to their higher production and lower costs of the final product. Modifications in the use of promoters, the use of recombination regions, and the change in the order of the chains, are some of the genetic engineering changes that can increase the production of recombinant antibodies. In this work, three different promoters were tested: Prom A, hCMV, and EF1-a, for two different antibodies, one anti-TNFa and one anti-CD20+. Changes were made in the order of the chains H-L or L-H and one or two UCOE (ubiquitous chromatin opening element) sequences were also used to identify the combinations that provide the best transient and stable expression for the antibodies in the CHO-s cells. In our results, we observed that the use of the two UCOE regions, with L-H order is almost three times better for the expression of the two different antibodies, while the strength of the promoter is conditioned by the sequence of each expressed protein.
Subject(s)
Key words

Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Gene Expression / Promoter Regions, Genetic / Tumor Necrosis Factor-alpha / Antigens, CD20 / Antibodies, Monoclonal Type of study: Prognostic_studies Limits: Animals Language: En Journal: Protein Expr Purif Journal subject: BIOLOGIA MOLECULAR Year: 2017 Document type: Article Affiliation country: México Country of publication: Estados Unidos

Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Gene Expression / Promoter Regions, Genetic / Tumor Necrosis Factor-alpha / Antigens, CD20 / Antibodies, Monoclonal Type of study: Prognostic_studies Limits: Animals Language: En Journal: Protein Expr Purif Journal subject: BIOLOGIA MOLECULAR Year: 2017 Document type: Article Affiliation country: México Country of publication: Estados Unidos