Production and N-glycan engineering of Varlilumab in Nicotiana benthamiana

Nguyen, Kim Dua; Kajiura, Hiroyuki; Kamiya, Ryo; Yoshida, Takahiro; Misaki, Ryo; Fujiyama, Kazuhito

doi:https://doi.org/10.3389/fpls.2023.1215580

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

アクセス数:468件（2025-07-18 10:12 集計）

固定URL: https://hdl.handle.net/11094/92492

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タイトル	Production and N-glycan engineering of Varlilumab in Nicotiana benthamiana
著者	著者名 Nguyen, Kim Dua
	NRID 1000030704209 1000030704209 ORCID 0000-0001-5811-1625 0000-0001-5811-1625 researchmap permalink 7000020437 7000020437 Scopus Author ID 16241653300 16241653300 著者名 Kajiura, Hiroyuki
	著者名 Kamiya, Ryo
	著者名 Yoshida, Takahiro
	NRID 1000020571186 1000020571186 researchmap permalink read0090369 read0090369 Scopus Author ID 6603301567 6603301567 著者名 Misaki, Ryo
	NRID 1000070209112 1000070209112 researchmap permalink read0014155 read0014155 Scopus Author ID 7005164172 7005164172 著者名 Fujiyama, Kazuhito
キーワード等	cancer immunotherapy
	galactosyltransferase
	hydroponics
	monoclonal antibody (mAb)
	Nicotiana benthamiana
	Varlilumab
内容	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.
抄録	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.
出版者	Frontiers Media SA
収録物名	Frontiers in Plant Science
巻(号)	14
ページ	1215580
刊行年月	2023-08-08
言語	英語
ハンドルURL	https://hdl.handle.net/11094/92492
eISSN	1664462X
関連情報 (isIdenticalTo)	DOI (出版社版) https://doi.org/10.3389/fpls.2023.1215580
アクセス権	open access
権利情報	© 2023 Nguyen, Kajiura, Kamiya, Yoshida, Misaki and Fujiyama. This article is licensed under a Creative Commons Attribution 4.0 International License.
出版タイプ	Version of Record
カテゴリ	学術雑誌論文 Journal Article

資源タイプ	学術雑誌論文
ローカル資源タイプ	学術雑誌論文
DCMI資源タイプ	text
DCTERMS.bibliographicCitation	Frontiers in Plant Science. 2023, 14, p. 1215580
DC.title	Production and N-glycan engineering of Varlilumab in Nicotiana benthamiana
DC.creator	Nguyen, Kim Dua
	Kajiura, Hiroyuki
	Kamiya, Ryo
	Yoshida, Takahiro
	Misaki, Ryo
	Fujiyama, Kazuhito
DC.publisher	Frontiers Media SA
DC.language' scheme='DCTERMS.RFC1766	英語
DC.type' scheme='DCTERMS.DCMIType	text
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2023-08-08
DC.identifier	https://doi.org/10.3389/fpls.2023.1215580
DC.subject	cancer immunotherapy
	galactosyltransferase
	hydroponics
	monoclonal antibody (mAb)
	Nicotiana benthamiana
	Varlilumab
DC.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.
DCTERMS.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.
DC.rights	© 2023 Nguyen, Kajiura, Kamiya, Yoshida, Misaki and Fujiyama. This article is licensed under a Creative Commons Attribution 4.0 International License.
DC.format	92492
citation_title	Production and N-glycan engineering of Varlilumab in Nicotiana benthamiana
citation_author	Nguyen, Kim Dua
	Kajiura, Hiroyuki
	Kamiya, Ryo
	Yoshida, Takahiro
	Misaki, Ryo
	Fujiyama, Kazuhito
citation_publisher	Frontiers Media SA
citation_language	英語
citation_date	2023-08-08
citation_journal_title	Frontiers in Plant Science
citation_volume	14
citation_firstpage	1215580
citation_doi	https://doi.org/10.3389/fpls.2023.1215580
citation_public_url	https://hdl.handle.net/11094/92492

Production and N-glycan engineering of Varlilumab in Nicotiana benthamiana

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