Conservation of receptor antagonist anti-tumor activity by epidermal growth factor receptor antibody expressed in transgenic corn seed

Ludwig, Dale L.; Witte, Larry; Hicklin, Daniel J.; Prewett, Marie; Bassi, Rajiv; Burtrum, Douglas; Pereira, Daniel S.; Jimenez, Xenia; Fox, Floyd E.; Saxena, B.; Zhou, Qinwei; Ma, Yuemei; Kang, Xiaoqiang; Patel, Dhaval; Barry, Michael A.; Kussie, Paul; Zhu, Zhenping; Russell, Debra; Petersen, William L.; Jury, Thomas P.; Gaitan-Gaitan, Fernando; Moran, Daniel L.; Delannay, Xavier; Storrs, Bradley S.; Tou, Jacob S.; Zupec, Mark E.; Gustafson, Karen S.; McIntyre, John B.; Tarnowski, S.Joseph; Bőhlen, Peter

doi:10.3233/hab-2004-13304

Cited by 16 publications

(6 citation statements)

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“…To restrict the recombinant protein to the endosperm, the rice gt-1 promoter was used, whose specificity has been confirmed in several studies (Russell and Fromm, 1997;Ludwig et al, 2004;Law et al, 2006). The identification of plant lines strongly expressing a recombinant protein is more challenging when using a seed-specific promoter, as expression levels can be analysed only after seed set.…”

Section: Discussionmentioning

confidence: 99%

“…Plants are being developed as inexpensive, large‐scale production systems to meet the future demand for complex biopharmaceuticals (Daniell et al ., 2001; Ma et al ., 2005; Schillberg et al ., 2005). Numerous studies have shown that transgenic plants can express correctly folded and functional antibodies, including full‐size immunoglobulin G (IgG) and secretory immunoglobulin A (sIgA) molecules (Ludwig et al ., 2004). Plant systems could therefore be ideal for the production of molecules such as 2G12, a human monoclonal antibody that recognizes oligomannose‐type N ‐glycans on the human immunodeficiency virus‐1 (HIV‐1) gp120 envelope protein.…”

Section: Introductionmentioning

confidence: 99%

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Recombinant antibody 2G12 produced in maize endosperm efficiently neutralizes HIV‐1 and contains predominantly single‐GlcNAc N‐glycans

Rademacher

Sack

Arcalís

et al. 2007

Plant Biotechnology Journal

166

132

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SummaryAntibody 2G12 is one of a small number of human immunoglobulin G (IgG) monoclonal antibodies exhibiting potent and broad human immunodeficiency virus-1 (HIV-1)-neutralizing activity in vitro , and the ability to prevent HIV-1 infection in animal models.It could be used to treat or prevent HIV-1 infection in humans, although to be effective it would need to be produced on a very large scale. We have therefore expressed this antibody in maize, which could facilitate inexpensive, large-scale production. The antibody was expressed in the endosperm, together with the fluorescent marker protein Discosoma red fluorescent protein (DsRed), which helps to identify antibody-expressing lines and trace transgenic offspring when bred into elite maize germplasm. To achieve accumulation in storage organelles derived from the endomembrane system, a KDEL signal was added to both antibody chains. Immunofluorescence and electron microscopy confirmed the accumulation of the antibody in zein bodies that bud from the endoplasmic reticulum. In agreement with this localization, N -glycans attached to the heavy chain were mostly devoid of Golgi-specific modifications, such as fucose and xylose. Surprisingly, most of the glycans were trimmed extensively, indicating that a significant endoglycanase activity was present in maize endosperm. The specific antigen-binding function of the purified antibody was verified by surface plasmon resonance analysis, and in vitro cell assays demonstrated that the HIV-neutralizing properties of the maize-produced antibody were equivalent to or better than those of its Chinese hamster ovary cell-derived counterpart.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Recombinant antibody 2G12 produced in maize endosperm efficiently neutralizes HIV‐1 and contains predominantly single‐GlcNAc N‐glycans

Rademacher

Sack

Arcalís

et al. 2007

Plant Biotechnology Journal

166

132

View full text Add to dashboard Cite

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“…[36][37][38][39] IGF-IR function is implicated in the hallmarks of cancer -self-sufficiency in growth signals, evasion from apoptosis, tissue invasion and metastasis, as well as angiogenesis. While we used ribozymes in this study, many other approaches have been used to study IGF-IR function including dominantnegative mutants, kinase-defective mutants, antisense oligonucleotides, antisense expression plasmids, IGFbinding proteins, soluble forms of the receptor, antagonistic and/or neutralizing antibodies [40][41][42][43][44] or small molecule kinase inhibitors. 45,46 These studies demonstrated that, in a variety of experimental settings, interference with IGF-IR function results in inhibition of cancer cell proliferation, 47 survival, 32 anchorageindependent growth in vitro, 48 inhibition of tumor growth and formation of metastasis in vivo 36,41,43,44,49 and sensitization of cancer cells to various chemotherapeutic and radiation regimens.…”

Section: Figurementioning

confidence: 99%

Proliferating endothelial cell-specific expression of IGF-I receptor ribozyme inhibits retinal neovascularization

et al. 2006

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“…These results are in line with those reported by Ludwig et al . (), which compared the anti‐EGFR chimeric antibody cetuximab, produced in mammalian cells, with glycosylated and aglycosylated antibody forms expressed in transgenic corn. These authors found that plantibodies were functionally indistinguishable from the mammalian cell–derived antibody in terms of binding to the receptor, the blockade of ligand‐dependent signalling and the inhibition of cell proliferation.…”

Section: Discussionmentioning

confidence: 99%

Comparative in vitro and experimental in vivo studies of the anti–epidermal growth factor receptor antibody nimotuzumab and its aglycosylated form produced in transgenic tobacco plants

Rodríguez

Pérez

Gavilondo

et al. 2012

Plant Biotechnology Journal

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SummaryA broad variety of foreign genes can be expressed in transgenic plants, which offer the opportunity for large-scale production of pharmaceutical proteins, such as therapeutic antibodies. Nimotuzumab is a humanized anti-epidermal growth factor receptor (EGFR) recombinant IgG1 antibody approved in different countries for the treatment of head and neck squamous cell carcinoma, paediatric and adult glioma, and nasopharyngeal and oesophageal cancers. Because the antitumour mechanism of nimotuzumab is mainly attributed to its ability to interrupt the signal transduction cascade triggered by EGF/EGFR interaction, we have hypothesized that an aglycosylated form of this antibody, produced by mutating the N 297 position in the IgG 1 Fc region gene, would have similar biochemical and biological properties as the mammalian-cell-produced glycosylated counterpart. In this paper, we report the production and characterization of an aglycosylated form of nimotuzumab in transgenic tobacco plants. The comparison of the plantibody and nimotuzumab in terms of recognition of human EGFR, effect on tyrosine phosphorylation and proliferation in cells in response to EGF, competition with radiolabelled EGF for EGFR, affinity measurements of Fab fragments, pharmacokinetic and biodistribution behaviours in rats and antitumour effects in nude mice bearing human A431 tumours showed that both antibody forms have very similar in vitro and in vivo properties. Our results support the idea that the production of aglycosylated forms of some therapeutic antibodies in transgenic plants is a feasible approach when facing scaling strategies for anticancer immunoglobulins.

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Conservation of receptor antagonist anti-tumor activity by epidermal growth factor receptor antibody expressed in transgenic corn seed

Cited by 16 publications

References 0 publications

Recombinant antibody 2G12 produced in maize endosperm efficiently neutralizes HIV‐1 and contains predominantly single‐GlcNAc N‐glycans

Recombinant antibody 2G12 produced in maize endosperm efficiently neutralizes HIV‐1 and contains predominantly single‐GlcNAc N‐glycans

Proliferating endothelial cell-specific expression of IGF-I receptor ribozyme inhibits retinal neovascularization

Comparative in vitro and experimental in vivo studies of the anti–epidermal growth factor receptor antibody nimotuzumab and its aglycosylated form produced in transgenic tobacco plants

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