Metabolism and metabolic burden by α1-antitrypsin production in human AGE1.HN cells

The metabolic burden on human AGE1.HN cells imposed by the production of recombinant α1-antitrypsin (A1AT) was studied by comparing a selected high-producing clonal cell line with the parental cell line. RNA, lipid, and phosphatidylcholine fractions were higher in the producer cell line causing metabolic changes in the producer, e.g., increased glycine and glutamate production. By simulating the theoretical metabolite demand for production of mature A1AT using a network model, it was found that the differences in metabolic profiles between producer and parental cells match the observed increased C1-unit and nucleotide demand as well as lipid precursor demand in the producer. Additionally, metabolic flux analysis revealed similar energy metabolism in both cell lines. The increased lipid content seems related to activated secretion machinery in the producer cell line. Increased lipid and C1 metabolism seem important targets for further improvement of AGE1.HN and other producing mammalian cells.

► New human cell line AGE1.HN produces fully human type α1-antitrypsin.
► Higher RNA and lipid content is typical for production and secretion of human α1-antitrypsin.
► In silico network analysis and metabolic flux analysis point to the same targets for improvement.
► Identified metabolic targets for α1-antitrypsin production are mainly C1 and lipid metabolism.
► This may be implemented by genetic engineering or by applying useful feeding strategies.