TY  - JOUR
AB  - Mammalian cells are commonly used to produce recombinant protein therapeutics, but suffer from a high cost per mg of protein produced. There is therefore great interest in improving protein yields to reduce production cost. We present an entirely novel approach to reach this goal through direct engineering of the cellular translation machinery by introducing the R98S point mutation in the catalytically essential ribosomal protein L10 (RPL10-R98S). Our data support that RPL10-R98S enhances translation levels and fidelity and reduces proteasomal activity in lymphoid Ba/F3 and Jurkat cell models. In HEK293T cells cultured in chemically defined medium, knock-in of RPL10-R98S was associated with a 1.7- to 2.5-fold increased production of four transiently expressed recombinant proteins and 1.7-fold for one out of two stably expressed proteins. In CHO-S cells, eGFP reached a 2-fold increased expression under stable but not transient conditions, but there was no production benefit for monoclonal antibodies. The RPL10-R98S associated production gain thus depends on culture conditions, cell type, and the nature of the expressed protein. Our study demonstrates the potential for using a ribosomal protein mutation for pharmaceutical protein production gains, and further research on how various factors influence RPL10-R98S phenotypes can maximize its exploitability for the mammalian protein production industry.
AU  - Verbelen, B.*
AU  - Girardi, T.*
AU  - Sulima, S.O.
AU  - Vereecke, S.*
AU  - Verstraete, P.*
AU  - Verbeeck, J.*
AU  - Royaert, J.*
AU  - Cinque, S.*
AU  - Montanaro, L.*
AU  - Penzo, M.*
AU  - Imbrechts, M.*
AU  - Geukens, N.*
AU  - Geuens, T.*
AU  - Dierckx, K.*
AU  - Pepe, D.*
AU  - Kampen, K.*
AU  - De Keersmaecker, K.*
C1  - 64098
C2  - 52079
CY  - 111 River St, Hoboken 07030-5774, Nj Usa
SP  - 100-114
TI  - Exploitation of the ribosomal protein L10 R98S mutation to enhance recombinant protein production in mammalian cells.
JO  - Eng. Life Sci.
VL  - 22
IS  - 2
PB  - Wiley
PY  - 2022
SN  - 1618-0240
ER  - 

TY  - JOUR
AB  - A new bacterial isolate (NH) from salt-affected soil was identified as Azospirillum brasilense using phenotypic analyses and 16SrDNA-based phylogeny. This isolate showed resistance towards 3,4-dehydroproline and optimal growth at 200 mmol/L NaCl, tolerating salt stress of 300 mmol/L NaCl in the absence of osmoprotectants and up to 600 mmol/L NaCl in the presence of glycine betaine and Ova lactuca extracts. This effect was enhanced with extracts of the marine algae Ova lactuca. A. brasilense strain NH can produce auxin indole acetic acid under saline conditions. The hypothesis was tested that the inoculation of this osmotolerant rhizosphere strain could improve the growth of wheat under saline stress conditions. Normal wheat growth was restored in the presence of both 150 mmol/L and 200 mmol/L NaCl after inoculation with A. brasilense NH. Under saline conditions, its effect of promoting plant growth of wheat was significantly superior to that of A. brasilense Sp7, the non-halotolerant type strain. A. brasilense NH restored wheat growth at elevated salt concentrations in pot and field experiments even better in the presence of osmoprotective Ulva lactuca extracts.
AU  - Nabti, E .*
AU  - Sahnoune, M.*
AU  - Adjrad, S.*
AU  - van Dommelen, A.
AU  - Ghoul, M.*
AU  - Schmid, M.
AU  - Hartmann, A.
C1  - 4169
C2  - 24834
SP  - 354-360
TI  - A halophilic and osmotolerant Azospirillum brasilense strain from Algerian soil restores wheat growth under saline conditions.
JO  - Eng. Life Sci.
VL  - 7
IS  - 4
PB  - Wiley-VCH
PY  - 2007
SN  - 1618-0240
ER  -