Inhibition of glutamine-dependent autophagy increases t-PA production in CHO cell fed-batch processes.
| Authors | Jardon MA, Sattha B, Braasch K, Leung AO, Côté HC, Butler M, Gorski SM & Piret JM. |
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| Abstract | Understanding the cellular responses caused by metabolic stress is crucial for the design of robust fed-batch bioprocesses that maximize the expression of recombinant proteins. Chinese hamster ovary cells were investigated in chemically defined, serum-free cultures yielding 10(7) cells/mL and up to 500 mg/L recombinant tissue-plasminogen activator (t-PA). Upon glutamine depletion increased autophagosome formation and autophagic flux were observed, along with decreased proliferation and high viability. Higher lysosomal levels correlated with decreased productivity. Chemical inhibition of autophagy with 3-methyl adenine (3-MA) increased the t-PA yield by 2.8-fold. Autophagy-related MAP1LC3 and LAMP2 mRNA levels increased continuously in all cultures. Analysis of protein quality revealed that 3-MA treatment did not alter glycan antennarity while increasing fucosylation, galactosylation, and sialylation. Taken together, these findings indicate that inhibition of autophagy can considerably increase the yield of biotechnology fed-batch processes, without compromising the glycosylation capacity of cells. Monitoring or genetic engineering of autophagy provides novel avenues to improve the performance of cell culture-based recombinant protein production. |
| Journal Name and Citation | Biotechnol Bioeng. 2012 May;109(5):1228-38. |
| Date of Publication | 2012/05/01 |
| Publication Link | http://onlinelibrary.wiley.com/doi/10.1002/bit.24393/abstract;jsessionid=9ACA47E9EF03BD13F0D3093B51168ADD.d02t02 |