Glycoengineering of plant-produced Pembrolizumab enhances FcRn binding and extends serum half-life in mice
Issued Date
2025-12-01
Resource Type
eISSN
2215017X
Scopus ID
2-s2.0-105017446811
Journal Title
Biotechnology Reports
Volume
48
Rights Holder(s)
SCOPUS
Bibliographic Citation
Biotechnology Reports Vol.48 (2025)
Suggested Citation
Bulaon C.J.I., Jaratsittisin J., Rattanapisit K., Suwanchaikasem P., Guo S., Boonha K., Pitaksajjakul P., Simsom N., Limprasutr V., Phoolcharoen W. Glycoengineering of plant-produced Pembrolizumab enhances FcRn binding and extends serum half-life in mice. Biotechnology Reports Vol.48 (2025). doi:10.1016/j.btre.2025.e00927 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/112472
Title
Glycoengineering of plant-produced Pembrolizumab enhances FcRn binding and extends serum half-life in mice
Author's Affiliation
Corresponding Author(s)
Other Contributor(s)
Abstract
Plant systems offer scalable and cost-effective platforms for antibody production, but plant-specific glycans may affect pharmacokinetics and immunogenicity. To evaluate the impact of Fc glycosylation, four Pembrolizumab glycovariants were generated in Nicotiana benthamiana: wild-type glycosylation (Pembro-WT), high-mannose with SEKDEL (Pembro-KD), aglycosylated N297A mutant (Pembro-NG), and a core fucose/xylose-deficient variant (Pembro-XF). Glycoproteins were transiently expressed either in wild-type or ΔXF plants, purified, and characterized for glycan composition, in vitro binding, and in vivo pharmacokinetics. LC-MS confirmed distinct glycoform patterns, while PD-1 binding was retained across all variants. Pembro-XF showed the highest FcRn binding affinity and longest serum half-life (45.83 h) in mice, compared to Pembro-WT (26.7 h), Pembro-KD (32.95 h), Pembro-NG (34.27 h), and Keytruda® (33.26 h). As an initial efficacy evaluation, Pembro-WT demonstrated strong antitumor activity in a murine colon cancer model. These findings support plant glycoengineering as a strategy to enhance antibody pharmacokinetics and advance next generation antibody therapeutics.