The C-terminal KDEL sequence increases the expression level of a single-chain antibody designed to be targeted to both the cytosol and the secretory pathway in transgenic tobacco

Schouten, Alexander; Roosien, J.; Engelen, F.A. van; Jong, G.A.M. de; Borst-Vrenssen, A.W.M.; Zilverentant, J.F.; Bosch, Dirk; Stiekema, Willem J.; Gommers, F.J.; Schots, A.; Bakker, J.

doi:10.1007/bf00019011

Cited by 258 publications

(144 citation statements)

References 38 publications

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“…That might be because of the use of strong 35S promoter containing a duplicated upstream enhancer region (18) to drive the expression of both HC and LC genes and͞or insertion of nontranslated viral leaders (tobacco etch virus and alfalfa mosaic virus) in front of the corresponding coding sequences. In addition, the ER retention signal KDEL (Lys-Asp-Glu-Leu) peptide was fused at the C terminus of the HC to enhance stability and increase overall accumulation of the protein (17,20).…”

Section: Discussionmentioning

confidence: 99%

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Plant-derived anti-Lewis Y mAb exhibits biological activities for efficient immunotherapy against human cancer cells

Brodzik

Glogowska²,

Bandurska³

et al. 2006

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

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Although current demands for therapeutic mAbs are growing quickly, production methods to date, including in vitro mammalian tissue culture and transgenic animals, provide only limited quantities at high cost. Several tumor-associated antigens in tumor cells have been identified as targets for therapeutic mAbs. Here we describe the production of mAb BR55-2 (IgG2a) in transgenic plants that recognizes the nonprotein tumor-associated antigen Lewis Y oligosaccharide overexpressed in human carcinomas, particularly breast and colorectal cancers. Heavy and light chains of mAb BR55-2 were expressed separately and assembled in plant cells of low-alkaloid tobacco transgenic plants (Nicotiana tabacum cv. LAMD609). Expression levels of plant-derived mAb (mAb P ) were high (30 mg͞kg of fresh leaves) in T 1 generation plants. Like the mammalian-derived mAb M , the plant mAb P bound specifically to both SK-BR3 breast cancer cells and SW948 colorectal cancer cells. The Fc domain of both mAb P and mAb M showed the similar binding to Fc␥RI receptor (CD64). Comparable levels of cytotoxicity against SK-BR3 cells were also shown for both mAbs in antibody-dependent cell-mediated cytotoxicity assay. Furthermore, plant-derived BR55-2 efficiently inhibited SW948 tumor growth xenografted in nude mice. Altogether, these findings suggest that mAb P originating from low-alkaloid tobacco exhibit biological activities suitable for efficient immunotherapy.breast and colorectal cancer ͉ plant biotechnology ͉ transgenic low-alkaloid tobacco ͉ tumor growth inhibition

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Section: Discussionmentioning

confidence: 99%

“…The mAb fusion to the KDEL signal sequence helped to retain the protein inside the endoplasmic reticulum (ER), thus enhancing mAb assembly in plant cells (17). Consequently, it helped to increase the final mAb yields from the plant production system.…”

mentioning

confidence: 99%

Plant-derived anti-Lewis Y mAb exhibits biological activities for efficient immunotherapy against human cancer cells

Brodzik

Glogowska²,

Bandurska³

et al. 2006

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

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“…Retention in the ER can be easily induced by adding retention signals KDEL or HDEL. In fact, ERretention has been repeatedly reported to enhance accumulation of recombinant antibodies in plants, most especially in the case of antibody fragments highly dependent on disulfide bridges for proper folding [56]. For certain proteins, ER milieu seems to provide a certain degree of stability, which is lost once the protein is delivered to the Golgi apparatus towards secretion.…”

Section: Two Edges Sword: Delivery Versus Disposalmentioning

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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“…Genes, those originally designed for the bacterial system is simply got modified to shuttle in the plant system by the addition of legumin signal peptide 35 . To improve the intracellular stability and yield percentage KDEL motif is incorporated into the sequence, which translocates the sequence into ER 24,36,37 . Irrespective of tissues constitutive promoters transcribe the genes effectively; this may be either good enough for monocotyledon and dicotyledonous plants.…”

Section: Construction Of Plant Expression Vectormentioning

confidence: 99%

Rapid production of therapeutic proteins using plant system

Ramalingam¹,

Iyappan²,

Priya³

et al. 2017

Def. Life. Sc. Jl.

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Plant molecular farming is simply defined as the production of proteins therapeutics (PT) in plants, which involves transient gene expression in plants and purification of expressed protein to a great scale for diagnosis, treatment and other applications. This is the rapid, economical, safe and reproducible approach for the production of PT as compared to bacterial and mammalian systems. Protein yield and post-translational modifications are the major roadblocks that can be overcome by high expression strategies includes over expression constructs, suitable plant host systems and glycoengineering of proteins. The inherent ability of ideally producing safe, functional protein is the most striking phenomenon recognised by the pharmaceutical industries and developed many therapeutic products within few weeks to meet escalating demands during pandemic/epidemic outbreaks recently.

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The C-terminal KDEL sequence increases the expression level of a single-chain antibody designed to be targeted to both the cytosol and the secretory pathway in transgenic tobacco

Cited by 258 publications

References 38 publications

Plant-derived anti-Lewis Y mAb exhibits biological activities for efficient immunotherapy against human cancer cells

Plant-derived anti-Lewis Y mAb exhibits biological activities for efficient immunotherapy against human cancer cells

Manufacturing antibodies in the plant cell

Rapid production of therapeutic proteins using plant system

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