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Glycan optimization of a human monoclonal antibody in the aquatic plant Lemna minor

Nature Biotechnology volume 24, pages 1591–1597 (2006)Cite this article

Abstract

N-glycosylation is critical to the function of monoclonal antibodies (mAbs) and distinguishes various systems used for their production. We expressed human mAbs in the small aquatic plant Lemna minor, which offers several advantages for manufacturing therapeutic proteins free of zoonotic pathogens1. Glycosylation of a mAb against human CD30 was optimized by co-expressing the heavy and light chains of the mAb with an RNA interference construct targeting expression of the endogenous α-1,3-fucosyltransferase and β-1,2-xylosyltransferase genes. The resultant mAbs contained a single major N-glycan species without detectable plant-specific N-glycans and had better antibody-dependent cell-mediated cytotoxicity and effector cell receptor binding activities than mAbs expressed in cultured Chinese hamster ovary (CHO) cells.

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Figure 1: RNAi silencing of α-1,3-fucosyltransferase and β-1,2-xylosyltransferase activities and analysis of purified MDX-060 from L. minor.
Figure 2: Mass spectrometric analysis of N-glycans labeled with 2-AA released from MDX-060 mAbs expressed in CHO (MDX-060 CHO), wild-type L. minor (MDX-060 LEX) or L. minor transformed with the Fuct1/Xylt1 RNAi construct (MDX-060 LEXOpt) grown at different scales (1 g or 300 g of L. minor tissue).
Figure 3: In vitro activities of MDX-060 mAbs.

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Acknowledgements

We wish to acknowledge Rachel Loranger, Betsy Adams, Heather Hilton, Vincent Wingate, Nirmala Rajbhandari, Lee Shirkey and Nancy Whitt for their work on the MDX-060 project and Ming-Bo Wang for his help in construct design.

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Authors and Affiliations

  1. Biolex Therapeutics, 158 Credle Street, Pittsboro, 27312, North Carolina, USA

    Kevin M Cox, Jason D Sterling, Jeffrey T Regan, John R Gasdaska, Karen K Frantz, Charles G Peele & Lynn F Dickey

  2. Medarex, Inc., 521 Cottonwood Drive, Milpitas, 95035, California, USA

    Amelia Black, David Passmore, Cristina Moldovan-Loomis, Mohan Srinivasan, Severino Cuison & Pina M Cardarelli

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Correspondence to Kevin M Cox.

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Supplementary information

Supplementary Table 1

Comparison of the thermal stabilities of MDX-060 CHO, MDX-060 LEX and glyco-optimized MDX-060 LEXOpt mAbs by differential scanning calorimetry (DSC) (DOC 24 kb)

Supplementary Table 2

Monosaccharides released from MDX-060 CHO, LEX and LEXOpt mAbs by acid hydrolysis and analyzed by HPAEC-PAD (DOC 26 kb)

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Cox, K., Sterling, J., Regan, J. et al. Glycan optimization of a human monoclonal antibody in the aquatic plant Lemna minor. Nat Biotechnol 24, 1591–1597 (2006). https://doi.org/10.1038/nbt1260

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