Production and N-glycan engineering of Varlilumab in Nicotiana benthamiana

Nguyen, Kim Dua; Kajiura, Hiroyuki; Kamiya, Ryo et al.

Frontiers in Plant Science, 2023, 14, 1215580

Number of Access:532（2025-10-24 17:54 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/92492

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Title
Production and N-glycan engineering of Varlilumab in Nicotiana benthamiana
Creator
CreatorName
Nguyen, Kim Dua
NRID
1000030704209 1000030704209
ORCID
0000-0001-5811-1625 0000-0001-5811-1625
researchmap permalink
7000020437 7000020437
Scopus Author ID
16241653300 16241653300
CreatorName
Kajiura, Hiroyuki
CreatorName
Kamiya, Ryo
CreatorName
Yoshida, Takahiro
NRID
1000020571186 1000020571186
researchmap permalink
read0090369 read0090369
Scopus Author ID
6603301567 6603301567
CreatorName
Misaki, Ryo
NRID
1000070209112 1000070209112
researchmap permalink
read0014155 read0014155
Scopus Author ID
7005164172 7005164172
CreatorName
Fujiyama, Kazuhito
Subject
cancer immunotherapy
galactosyltransferase
hydroponics
monoclonal antibody (mAb)
Nicotiana benthamiana
Varlilumab
Description
Nguyen K.D., Kajiura H., Kamiya R., et al. Production and N-glycan engineering of Varlilumab in Nicotiana benthamiana. Frontiers in Plant Science 14, 1215580 (2023); https://doi.org/10.3389/fpls.2023.1215580.
Abstract
N-glycan engineering has dramatically evolved for the development and quality control of recombinant antibodies. Fc region of IgG contains two N-glycans whose galactose terminals on Fc-glycan have been shown to increase the stability of CH2 domain and improve effector functions. Nicotiana benthamiana has become one of the most attractive production systems for therapeutic antibodies. In this study, Varlilumab, a CD27-targeting monoclonal antibody, was transiently produced in fresh leaves of soil-grown and hydroponic-grown N. benthamiana, resulted in the yield of 174 and 618 µg/gram, respectively. However, the IgG produced in wild-type N. benthamiana lacked the terminal galactose residues in its N-glycan. Therefore, N-glycan engineering was applied to fine-tune recombinant antibodies produced in plant platforms. We further co-expressed IgG together with murine β1,4-galactosyltransferase (β1,4-GALT) to modify plant N-glycan with β1,4-linked Gal residue(s) and Arabidopsis thaliana β1,3-galactosylatransferase (β1,3-GALT) to improve galactosylation. The co-expression of IgG with each of GALTs successfully resulted in modification of N-glycan structures on the plant-produced IgG. Notably, IgG co-expressed with murine β1,4-GALT in soil-grown N. benthamiana had 42.5% of N-glycans variants having galactose (Gal) residues at the non-reducing terminus and 55.3% of that in hydroponic-grown N. benthamiana plants. Concomitantly, N-glycan profile analysis of IgG co-expressed with β1,3-GALT demonstrated that there was an increased efficiency of galactosylation and an enhancement in the formation of Lewis a structure in plant-derived antibodies. Taken together, our findings show that the first plant-derived Varlilumab was successfully produced with biantennary β1,4-galactosylated N-glycan structures.
Publisher
Frontiers Media SA
Source Title
Frontiers in Plant Science
Volume (Issue)
14
Page
1215580
Date of Issued
2023-08-08
Language
English
Handle URL
https://hdl.handle.net/11094/92492
eISSN
1664462X
Relation.isIdenticalTo
DOI (Publisher Version)
https://doi.org/10.3389/fpls.2023.1215580
Access Rights
open access
CopyRight
© 2023 Nguyen, Kajiura, Kamiya, Yoshida, Misaki and Fujiyama. This article is licensed under a Creative Commons Attribution 4.0 International License.
oaire:version
Version of Record
Category
学術雑誌論文 Journal Article