AUTHOR=Sariyatun Ratna , Florence  , Kajiura Hiroyuki , Ohashi Takao , Misaki Ryo , Fujiyama Kazuhito 

TITLE=Production of Human Acid-Alpha Glucosidase With a Paucimannose Structure by Glycoengineered Arabidopsis Cell Culture

JOURNAL=Frontiers in Plant Science

VOLUME=12

YEAR=2021

URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2021.703020

DOI=10.3389/fpls.2021.703020

ISSN=1664-462X

ABSTRACT=<p>Plant cell cultures have emerged as a promising platform for the production of biopharmaceutics due to their cost-effectiveness, safety, ability to control the cultivation, and secrete products into culture medium. However, the use of this platform is hindered by the generation of plant-specific <italic>N</italic>-glycans, the inability to produce essential <italic>N</italic>-glycans for cellular delivery of biopharmaceutics, and low productivity. In this study, an alternative acid-alpha glucosidase (GAA) for enzyme replacement therapy of Pompe disease was produced in a glycoengineered <italic>Arabidopsis alg3</italic> cell culture. The <italic>N</italic>-glycan composition of the GAA consisted of a predominantly paucimannosidic structure, Man<sub>3</sub>GlcNAc<sub>2</sub> (M3), without the plant-specific <italic>N</italic>-glycans. Supplementing the culture medium with NaCl to a final concentration of 50 mM successfully increased GAA production by 3.8-fold. GAA from an NaCl-supplemented culture showed a similar <italic>N</italic>-glycan profile, indicating that the NaCl supplementation did not affect <italic>N</italic>-glycosylation. The results of this study highlight the feasibility of using a glycoengineered plant cell culture to produce recombinant proteins for which M3 or mannose receptor-mediated delivery is desired.</p>