Cost-effective production of a vaginal protein microbicide to prevent HIV transmission

Ramessar, Koreen; Rademacher, Thomas W.; Sack, Markus; Stadlmann, Johannes; Platis, Dimitris; Stiegler, Gabriela; Labrou, Nikolaos E.; Altmann, Friedrich; Ma, Julian; Stöger, Eva; Capell, Teresa; Christou, Paul

doi:10.1073/pnas.0708841104

Cited by 148 publications

(133 citation statements)

References 49 publications

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“…Thirteen-dayold immature zygotic embryos were bombarded with 10 mg of gold particles coated with the carotenogenic constructs and the selectable marker bar (41) at a molar ratio of 3:1 as reported (42) and returned to osmoticum medium for 12 h before selection. Bombarded callus was selected on phosphinothricinsupplemented medium as described (43,44), and transgenic plantlets were regenerated and hardened off in soil. Seventy independent events were selected for further analysis and either pollinated with nontransformed white maize (M37W) pollen or self-pollinated to produce T1 seeds.…”

Section: Discussionmentioning

confidence: 99%

Combinatorial genetic transformation generates a library of metabolic phenotypes for the carotenoid pathway in maize

Zhu

Naqvi

Breitenbach

et al. 2008

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

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Combinatorial nuclear transformation is a novel method for the rapid production of multiplex-transgenic plants, which we have used to dissect and modify a complex metabolic pathway. To demonstrate the principle, we transferred 5 carotenogenic genes controlled by different endosperm-specific promoters into a white maize variety deficient for endosperm carotenoid synthesis. We recovered a diverse population of transgenic plants expressing different enzyme combinations and showing distinct metabolic phenotypes that allowed us to identify and complement rate-limiting steps in the pathway and to demonstrate competition between ␤-carotene hydroxylase and bacterial ␤-carotene ketolase for substrates in 4 sequential steps of the extended pathway. Importantly, this process allowed us to generate plants with extraordinary levels of ␤-carotene and other carotenoids, including complex mixtures of hydroxycarotenoids and ketocarotenoids. Combinatorial transformation is a versatile approach that could be used to modify any metabolic pathway and pathways controlling other biochemical, physiological, or developmental processes.induced mutation ͉ metabolic engineering ͉ transgenic plant ͉ provitamin A

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Section: Discussionmentioning

confidence: 99%

Combinatorial genetic transformation generates a library of metabolic phenotypes for the carotenoid pathway in maize

Zhu

Naqvi

Breitenbach

et al. 2008

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

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328

263

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“…Immature zygotic embryos of the South African élite white maize M37W inbred were transformed by particle bombardment according to Ramessar et al (2008). Bombarded embryos were incubated on N6-based médium under dark for 15 days and then transferred every 2 weeks to fresh N6-based médium containing 3 mg/1 phosphinothricin (PPT).…”

Section: Plasmid Constructs and Plant Transformationmentioning

confidence: 99%

Expression of a barley cystatin gene in maize enhances resistance against phytophagous mites by altering their cysteine-proteases

et al. 2010

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Phytocystatins are inhibitors of cysteine-proteases from plants putatively involved in plant defence based on their capability of inhibit heterologous enzymes. We have previously characterised the whole cystatin gene family members from barley (HvCPI-1 to HvCPI-13). The aim of this study was to assess the effects of barley cystatins on two phytophagous spider mites, Tetranychus urticae and Brevipalpus chilensis. The determination of proteolytic activity profile in both mite species showed the presence of the cysteine-proteases, putative targets of cystatins, among other enzymatic activities. All barley cystatins, except HvCPI-1 and HvCPI-7, inhibited in vitro mite cathepsin L-and/or cathepsin B-like activities, HvCPI-6 being the strongest inhibitor for both mite species. Transgenic maize plants expressing HvCPI-6 protein were generated and the functional integrity of the cystatin transgene was confirmed by in vitro inhibitory effect observed against T urticae and B. chilensis protein extracts. Feeding experiments impaired on transgenic lines performed with T urticae impaired mite development and reproductive performance. Besides, a significant reduction of cathepsin L-like and/or cathepsin B-like activities was observed when the spider mite fed on maize plants expressing HvCPI-6 cystatin. These findings reveal the potential of barley cystatins as acaricide proteins to protect plants against two important mite pests.

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“…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.…”

Section: Full Size Antibodiesmentioning

confidence: 99%

Manufacturing antibodies in the plant cell

Orzáez

Granell

Blázquez

2009

Biotechnology Journal

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International audiencePlants have long been considered advantageous platforms for large-scale production of antibodies due to their low cost, scalability, and the low chances of pathogen contamination. Much effort has therefore been devoted to efficiently produce mAbs (from nanobodies to secretory antibodies) in plant cells. Several technical difficulties have been encountered and are being coped with. Improvements in production levels have been achieved by manipulation of gene expression and, more efficiently, of cell targeting and protein folding and assembly. Differences in mAb glycosylation patterns between animal and plant cells are being successfully addressed by the elimination and introduction of the appropriate enzyme activities in plant cells. Another relevant battlefield is the dichotomy between production capacity and speed. Classically, stably transformed plant lines have been proposed for large scale mAb production, whereas the use of transient expression systems has always provided production speed at the cost of scalability. However, recent advances in transient expression techniques have brought impressive yield improvements, turning speed and scalability highly compatible assets. In the era of personalized medicines, the combination of yield and speed, and the advances in glyco-engineering have made the plant cell a serious contender in the field of recombinant antibody production

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Cost-effective production of a vaginal protein microbicide to prevent HIV transmission

Cited by 148 publications

References 49 publications

Combinatorial genetic transformation generates a library of metabolic phenotypes for the carotenoid pathway in maize

Combinatorial genetic transformation generates a library of metabolic phenotypes for the carotenoid pathway in maize

Expression of a barley cystatin gene in maize enhances resistance against phytophagous mites by altering their cysteine-proteases

Manufacturing antibodies in the plant cell

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