Production of antibodies in transgenic plants

Abstract

Complementary DNAs derived from a mouse hybridoma messenger RNA were used to transform tobacco leaf segments followed by regeneration of mature plants. Plants expressing single gamma or kappa immunoglobulin chains were crossed to yield progeny in which both chains were expressed simultaneously. A functional antibody accumulated to 1.3% of total leaf protein in plants expressing full-length cDNAs containing leader sequences. Specific binding of the antigen recognized by these antibodies was similar to the hybridoma-derived antibody. Transformants having gamma- or kappa-chain cDNAs without leader sequences gave poor expression of the proteins. The increased abundance of both gamma- and kappa-chains in transformants expressing assembled gamma-kappa complexes was not reflected in increased mRNA levels. The results demonstrate that production of immunoglobulins and assembly of functional antibodies occurs very efficiently in tobacco. Assembly of subunits by sexual cross might be a generally applicable method for expression of heterologous multimers in plants.

Manufacturing antibodies in the plant cell

“…In consequence, the development phase for recombinant mAb production is accelerated. Nevertheless, form the pionner work of Hiatt et al [13] to the most recent production of anti-HIV prophylactic IgG in corn [21,22], the lengthy process of establishing a homozygous transgenic plant line with stable and homogeneous IgG production has been the Achilles' heel of plant-made antibodies.…”

“…Unlike single chain (scFv) antibody fragments, which could be produced as periplasmic proteins in bacterial fermentors, the more complex full-size recombinant antibodies required eukaryotic cells as biofactories. Plants, among others, were postulated from the beginning as a suitable production platform [13]. Transgenic plants' technology was however too slow and rudimentary to compete, and CHO cells finally became the standard for the antibody industry.…”

Les microalgues

“…De plus, les organismes photosynthétiques offrent deux voies biologiques très différentes dans l'expression et la production de molécules recombinantes : une voie nucléaire et une voie chloroplastique (Tableau I). Les travaux pionniers de Hiatt avec la production d'anticorps fonctionnels exprimés dans du tabac transgénique, ont consacré le terme de Plantibodies (ou planticorps) [7]. De nombreuses autres molécules recombinantes actives, vaccins, anticorps monoclonaux, enzymes et hormones, ont été depuis produites avec succès [8].…”

Production of recombinant single chain antibodies (scFv) in vegetatively reproductive Kalanchoe pinnata by in planta transformation

“…Plant system has several advantages over other eukaryotes used for the production of recombinant proteins, namely: (1) the production cost of proteins from plant is less expensive than those from transgenic animals, fermentation, bioreactors, and microbial or animal cell culture-based systems; (2) plant systems can be scaled up quickly; (3) plants do not have any known human or mammalian pathogens; (4) plant cells are able to carry out all of the correct folding, glycosylation, and other posttranslational activities necessary for the accurate production of foreign proteins; and (5) plant cells can direct proteins to environments to reduce degradation and therefore increase stability (Horn et al 2004). Following the first report of successful production of a mouse monoclonal antibody in a plant (Hiatt et al 1989), many other therapeutic proteins have been expressed in transgenic plants. In general, four approaches have been considered for developing molecular farming system in crop species (Fischer et al 2004): nuclear transformation, plastid (chloroplast) transformation, virus-mediated transient transformation, and stable transformation of hydroponically-grown plant species.…”