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Enhancing the secretion of a glyco-engineered anti-CD20 scFv-Fc antibody in hairy root cultures
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Lonoce, Chiara, Marusic, Carla, Morrocchi, Elena, Salzano, Anna Maria, Scaloni, Andrea, Novelli, Flavia, Pioli, Claudio, Feeney, Mistianne, Frigerio, Lorenzo and Donini, Marcello (2019) Enhancing the secretion of a glyco-engineered anti-CD20 scFv-Fc antibody in hairy root cultures. Biotechnology Journal, 14 (3). 1800081. doi:10.1002/biot.201800081 ISSN 1860-6768.
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Official URL: https://doi.org/10.1002/biot.201800081
Abstract
Hairy root (HR) cultures represent an attractive platform for the production of heterologous proteins, due to the possibility of secreting the molecule of interest in the culture medium. The main limitation is the low accumulation yields of heterologous proteins. The aim of this study is to enhance the accumulation of a tumor‐targeting antibody with a human‐compatible glycosylation profile in HR culture medium. To this aim, the authors produce Nicotiana benthamiana HR cultures expressing the red fluorescent protein (RFP) to easily screen for different auxins able to induce heterologous protein secretion in the medium. The hormone 2,4‐dichlorophenoxyacetic acid (2,4‐D) is found to induce high accumulation levels (334 mg L−1) of RFP in the culture medium. The same protocol is used to improve the secretion of the tumor‐targeting, CD20‐specific 2B8‐FcΔXF recombinant antibody from glyco‐engineered ΔXTFT N. benthamiana HR cultures. The addition of 2,4‐D determine a 28‐fold increase of the accumulation of fully functional 2B8‐FcΔXF in the culture medium, at levels of ≈16 mg L−1. Antibody N‐glycosylation profiling reveal the prominent occurrence of GnGn structures and low levels of xylose‐ and fucose‐containing counterparts. This result is the first example of the expression of an engineered anti‐CD20 antibody with a scFv‐Fc format at high levels in HR.
Item Type: | Journal Article | ||||||||
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Divisions: | Faculty of Science, Engineering and Medicine > Science > Life Sciences (2010- ) | ||||||||
SWORD Depositor: | Library Publications Router | ||||||||
Journal or Publication Title: | Biotechnology Journal | ||||||||
Publisher: | Wiley-Blackwell Publishing Ltd. | ||||||||
ISSN: | 1860-6768 | ||||||||
Official Date: | March 2019 | ||||||||
Dates: |
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Volume: | 14 | ||||||||
Number: | 3 | ||||||||
Article Number: | 1800081 | ||||||||
DOI: | 10.1002/biot.201800081 | ||||||||
Status: | Peer Reviewed | ||||||||
Publication Status: | Published | ||||||||
Access rights to Published version: | Restricted or Subscription Access | ||||||||
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