Recombinant antibody 2G12 produced in maize endosperm efficiently neutralizes HIV-1 and contains predominantly single-GlcNAc N-glycans
Thomas W. Rademacher,Markus Sack,Elsa Arcalis,Johannes Stadlmann,Simone Balzer,Friedrich Altmann,Heribert Quendler,Gabriela Stiegler,Renate Kunert,Rainer Fischer,Eva Stoger +10 more
Reads0
Chats0
TLDR
In this paper, a maize-produced antibody was 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.Abstract:
Antibody 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.read more
Citations
More filters
Journal ArticleDOI
Plant-specific glycosylation patterns in the context of therapeutic protein production.
Véronique Gomord,Anne-Catherine Fitchette,Laurence Menu-Bouaouiche,Claude Saint-Jore-Dupas,Carole Plasson,Dominique Michaud,Loïc Faye +6 more
TL;DR: This review describes protein N- and O-linked glycosylation in plants and highlights the limitations and advantages of plant-specific gly cosylation on plant-made biopharmaceuticals.
Journal ArticleDOI
Analysis of immunoglobulin glycosylation by LC-ESI-MS of glycopeptides and oligosaccharides.
Johannes Stadlmann,Martin Pabst,Daniel Kolarich,Daniel Kolarich,Renate Kunert,Friedrich Altmann +5 more
TL;DR: They both outperform non‐MS methods in terms of reliability of peak assignment and MALDI‐MS of underivatized glycans with regard to the recording of sialylated structures and Regarding fast and yet detailed structural assignment, LC‐MS on graphitic carbon supersedes all other current methods.
Journal ArticleDOI
Extremely High-Level and Rapid Transient Protein Production in Plants without the Use of Viral Replication
TL;DR: A system based on a disabled version of cowpea mosaic virus RNA-2, which overcomes limitations on insert size and introduces biocontainment, provides an ideal vehicle for high-level expression that does not rely on viral replication of transcripts.
Journal ArticleDOI
Expression of recombinant Antibodies
TL;DR: This review focuses on current antibody production systems including their usability for different applications, and recent developments of glycosylation-engineered yeast, insect cell lines, and transgenic plants are promising to obtain antibodies with “human-like” post-translational modifications.
Journal ArticleDOI
Advances in plant molecular farming.
TL;DR: The various cost-effective technologies and strategies, which are being developed to improve yield and quality of the plant-derived pharmaceuticals, thereby making plant-based production system suitable alternatives to the existing systems are reviewed.
References
More filters
Journal ArticleDOI
A simple method of estimating fifty per cent endpoints
Lowell J. Reed,H. Muench +1 more
Journal ArticleDOI
Production of recombinant protein therapeutics in cultivated mammalian cells
TL;DR: Recently, the productivity of mammalian cells cultivated in bioreactors has reached the gram per liter range in a number of cases, a more than 100-fold yield improvement over titers seen for similar processes in the mid-1980s.
Journal ArticleDOI
Protection of macaques against vaginal transmission of a pathogenic HIV-1/SIV chimeric virus by passive infusion of neutralizing antibodies.
John R. Mascola,Gabriela Stiegler,Thomas C. VanCott,Hermann Katinger,Calvin B. Carpenter,Chris E. Hanson,Holly Beary,Deborah Hayes,Sarah S. Frankel,Deborah L. Birx,Mark G. Lewis +10 more
TL;DR: It is demonstrated that antibodies can affect transmission and subsequent disease course after vaginal SHIV-challenge, and the data begin to define the type of antibody response that could play a role in protection against mucosal transmission of HIV-1.
Journal ArticleDOI
Engineered glycoforms of an antineuroblastoma IgG1 with optimized antibody-dependent cellular cytotoxic activity
TL;DR: The glycosylation pattern of chCE7 was engineered in Chinese hamster ovary cells with tetracycline–regulated expression of GnTIII to optimize the ADCC activity, and this activity correlated with the level of constant region–associated, bisected complex oligosaccharides determined by matrix–assisted laser desorption/ionization time–of–flight mass spectrometry.