Monoclonal C5‐1 antibody produced in transgenic alfalfa plants exhibits a N‐glycosylation that is homogenous and suitable for glyco‐engineering into human‐compatible structures

Bardor, Muriel; Loutelier‐Bourhis, Corinne; Paccalet, Thomas; Cosette, Pascal; Fitchette, Anne‐Catherine; Vézina, Louis‐P.; Trépanier, Sonia; Dargis, Michèle; Lemieux, Réal; Lange, Catherine; Faye, Loı̈c; Lerouge, Patrice

doi:10.1046/j.1467-7652.2003.00041.x

Cited by 84 publications

(47 citation statements)

References 38 publications

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“…6) (22 (22). This glycosylation profile of mAb P CO17-1A differs from that in previous reports of other mAb P s (17,27,28). The profile discrepancy reflects the intrinsic properties of the Fc region of mAbs, which affect the accessibility of the individual glycans and inherent efficiency of the mAb secretion in plants (7).…”

Section: Discussioncontrasting

confidence: 62%

Inhibition of tumor growth by plant-derived mAb

Steplewski

Glogowska

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

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The tumor-associated antigen EpCAM (GA733-2) is a highly expressed target on adenocarcinoma cells, as defined by murine mAb CO17-1A. We recently developed a transgenic plant system for the safe and inexpensive production of large quantities of mAb CO17-1A as a future source of clinical-grade protein. Although the glycosylation pattern of plant-derived mAb (mAb P ) CO17-1A differs considerably from that of the mammalian-derived mAb (mAb M ), we show here that the biological activity of both mAbs is quite similar. mAb P heavy and light chains assembled to bind the recombinant antigen GA733-2E and specifically bound to human SW948 colorectal carcinoma cells expressing the antigen GA733-2 to the same extent as mAb M . mAb P was as effective as mAb M CO17-1A in inhibiting tumor growth of xenotransplanted SW948 cells in nude mice. These results suggest the promise of transgenic plants as a useful alternative way to produce full-size mAb for cancer immunotherapy.endoplasmic reticulum ͉ glycosylation ͉ Lys-Asp-Glu-Leu ͉ mammalianderived mAb

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Section: Discussioncontrasting

confidence: 62%

Inhibition of tumor growth by plant-derived mAb

Steplewski

Glogowska

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

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“…In Vitro Galactosylation-The in vitro galactosylation was performed by treating 50 g of protein at 37°C for 24 h with 50 milliunits of ␤(1,4)-galactosyltransferase from bovine milk in 1 ml of 100 mM sodium cacodylate buffer, pH 6.4, supplemented with 5 mol of UDP-galactose and 5 mol of MnCl 2 (34). The sample was then freeze-dried.…”

Section: Methodsmentioning

confidence: 99%

“…Proteins and glycoproteins were separated by SDS-PAGE and electroblotted onto nitrocellulose membrane. Glycoproteins were then affinodetected using biotinylated RCA 120 lectin (34).…”

Section: Methodsmentioning

confidence: 99%

“…To investigate the presence in P. tricornutum proteins of complex glycans carrying terminal GlcNAc, we treated the protein extract with a ␤(1,4)-galactosyltransferase, an enzyme able to transfer a galactose residue onto terminal GlcNAc residues, and then analyzed by affinoblotting the resulting protein preparation with RCA 120, a lectin that binds specifically to Gal␤1-4GlcNAc sequences (34). In contrast to a plant-derived IgG used as a positive control of galactose transfer, no signal was detected in P. tricornutum sample after this treatment (Fig.…”

Section: Journal Of Biological Chemistry 6157mentioning

confidence: 99%

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N-Glycans of Phaeodactylum tricornutum Diatom and Functional Characterization of Its N-Acetylglucosaminyltransferase I Enzyme

Baïet

Burel²,

Saint‐Jean³

et al. 2011

Journal of Biological Chemistry

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127

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N-Glycosylation, a major co-and post-translational event in the synthesis of proteins in eukaryotes, is unknown in aquatic photosynthetic microalgae. In this paper, we describe the Nglycosylation pathway in the diatom Phaeodactylum tricornutum. Bio-informatic analysis of its genome revealed the presence of a complete set of sequences potentially encoding for proteins involved in the synthesis of the lipid-linked Glc 3 Man 9 GlcNAc 2 -PP-dolichol N-glycan, some subunits of the oligosaccharyltransferase complex, as well as endoplasmic reticulum glucosidases and chaperones required for protein quality control and, finally, the ␣-mannosidase I involved in the trimming of the N-glycan precursor into Man-5 N-glycan. Moreover, one N-acetylglucosaminyltransferase I, a Golgi glycosyltransferase that initiates the synthesis of complex type N-glycans, was predicted in the P. tricornutum genome. We demonstrated that this gene encodes for an active N-acetylglucosaminyltransferase I, which is able to restore complex type N-glycans maturation in the Chinese hamster ovary Lec1 mutant, defective in its endogeneous N-acetylglucosaminyltransferase I. Consistent with these data, the structural analyses of N-linked glycans demonstrated that P. tricornutum proteins carry mainly high mannose type N-glycans ranging from Man-5 to Man-9. Although representing a minor glycan population, paucimannose N-glycans were also detected, suggesting the occurrence of an N-acetylglucosaminyltransferase I-dependent maturation of N-glycans in this diatom.

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“…6.5 ). In general, plant-derived proteins are a heterogeneous N-glycan group, with a few exceptions, e.g., the monoclonal C5-1 antibody produced in alfalfa (Bardor et al 2003 ;D'Aoust et al 2004 ).…”

Section: Plant Glycosylationmentioning

confidence: 99%

Molecular Farming in Plants

Álvarez

2014

Plant Biotechnology for Health

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Molecular farming can generally be defi ned as the production of molecules (proteins, fatty acids) for the pharmaceutical and chemical industries in transgenic organisms (plants, animals, etc.).Molecular farming in plants has advantageous aspects as biosecurity, they do not bear pathogens for humans or animals, and they do not produce toxins. Also, plant protein synthetic machinery is able to produce complex glycosylated proteins as they have a glycosylation pattern with slight difference respect of that of mammals, and can perform foldings.When displayed in in vitro conditions, molecular farming have the characteristic advantages of that type of culture, mainly the capacity of working under Good Manufactory practices as is required by the pharmaceutical industry. The critical aspects of plants for molecular farming are the different glycosylation patterns respect to mammals, the duration of the productive process, and the relatively low yields.The approval by the FDA of the fi rst medicine to be used in humans, taliglucerase alfa (ELELYSO®), has give an impulse to this technology but there are some drawbacks to be addressed, particularly related to low yields and regulatory aspects.

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Monoclonal C5‐1 antibody produced in transgenic alfalfa plants exhibits a N‐glycosylation that is homogenous and suitable for glyco‐engineering into human‐compatible structures

Cited by 84 publications

References 38 publications

Inhibition of tumor growth by plant-derived mAb

Inhibition of tumor growth by plant-derived mAb

N-Glycans of Phaeodactylum tricornutum Diatom and Functional Characterization of Its N-Acetylglucosaminyltransferase I Enzyme

Molecular Farming in Plants

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