The effect of the cell death suppressor vMIA on the production of a recombinant protein in the adenovirus-293 expression system.
Apoptosis is a major problem in animal cell cultures during production of biopharmaceuticals, such as recombinant proteins or viral vectors. A 293 cell line constitutively expressing vMIA (viral mitochondria-localized inhibitor of apoptosis) was constructed and examined on production of a model recombinant protein, green fluorescent protein (GFP) in the adenovirus-293 expression system, and on production of a model infectious adenoviral vector. vMIA-293 cells were more resistant than the parental 293 cells to apoptosis induced by either oxidative stress, or by adenovirus infection. The yield of GFP produced in vMIA-293 cell cultures was consistently higher (approximately 140%) compared to that in the parental cells. vMIA reduced production of adenovirus infectious particles, which was not due to a decline of adenovirus replication, since adenoviral DNA replication rate in vMIA-293 cells was higher than that in the parental cells. In conclusion, introduction of the vMIA gene into the 293 cell line is a promising strategy to improve recombinant protein production in the adenovirus-293 expression system.
Vieira
Helena L A
HL
Instituto de Tecnologia Química e Biológica (ITQB-UNL), Instituto de Biologia Experimental e Tecnológica (IBET), Apartado 12, 2781-901 Oeiras, Portugal.
Cunha
Liliana
L
Goldmacher
Victor S
VS
Alves
Paula M
PM
D000256:Adenoviridae / Q000235:genetics* / Q000378:metabolism; D017209:Apoptosis; D016923:Cell Death; D002478:Cells, Cultured; D005434:Flow Cytometry; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000235:genetics* / Q000378:metabolism; D018384:Oxidative Stress; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism
D024101:Mitochondrial Proteins; D011994:Recombinant Proteins; D014764:Viral Proteins; D049452:Green Fluorescent Proteins