Evaluation of changes in promoters, use of UCOES and chain order to improve the antibody production in CHO cells.
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.
Centro de Biotecnología-FEMSA, Tecnológico de Monterrey at Monterrey, Ave. Eugenio Garza Sada 2501 Sur Col. Tecnológico, CP 64849, Monterrey, Nuevo León, Mexico.
Maria Del Refugio
MDR
Rocha-Pizaña
Centro de Biotecnología-FEMSA, Tecnológico de Monterrey at Monterrey, Ave. Eugenio Garza Sada 2501 Sur Col. Tecnológico, CP 64849, Monterrey, Nuevo León, Mexico.
Guadalupe
G
Ascencio-Favela
Centro de Biotecnología-FEMSA, Tecnológico de Monterrey at Monterrey, Ave. Eugenio Garza Sada 2501 Sur Col. Tecnológico, CP 64849, Monterrey, Nuevo León, Mexico.
Brenda Maribell
BM
Soto-García
Molecular Medicine Laboratory, Unidad Academica de Medicina Humana y Ciencias de la Salud de la Universidad Autonoma de Zacatecas, Zacatecas, Mexico.
Margarita de la Luz
ML
Martinez-Fierro
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.
Mario Moisés
MM
Alvarez
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000235:genetics; D018951:Antigens, CD20; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D015870:Gene Expression; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014409:Tumor Necrosis Factor-alpha
D000911:Antibodies, Monoclonal; D018951:Antigens, CD20; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha
Antibody; CHO-s; EF1-a; L-H chains; UCOE; hCMV; promA