Expression of anti-sclerotinia scFv in transgenic Brassica napus enhances tolerance against stem rot

Yajima, William; Verma, Shiv; Shah, Saleh; Rahman, Masudur; Liang, Yue; Kav, Nat N. V.

doi:10.1016/j.nbt.2010.09.010

Cited by 23 publications

(22 citation statements)

References 19 publications

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“…First, one can employ antibody in miniaturized format based on the observation that H chains retain some antigen-binding capacity even in the absence of L chains. The smallest format found to keep full binding activity is the so-called single-chain fragment (scFv), in which the two variable regions, VH and VL, are artificially linked by a flexible polypeptide [12]–[15]. The other miniaturized type is the H-chain antibody derived from camelids (camels and llamas) where the L chain is missing [33].…”

Section: Discussionmentioning

confidence: 99%

“…These miniaturized antibodies do not require glycosylation and can be assembled in both plant and prokaryotic systems such as Escherichia coli . For example, scFv-based antibodies against HER2/neu have been produced in E. coli and in plants using both stable and transient systems in tobacco and Nicotiana benthamiana [12]–[15].…”

Section: Discussionmentioning

confidence: 99%

“…It has been estimated that the cost of pharmaceutical protein production in plants could be 10- to 50-fold lower than production of the same protein in mammals [10], [11]. Plants rapidly accumulate single-chain [12]–[15] and full-size antibodies [16]–[20] and may produce personalised patient-specific anticancer vaccines [21]. Plants may be a source of biosimilars, new versions of known pharmaceuticals, including anticancer antibodies [22].…”

Section: Introductionmentioning

confidence: 99%

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Plant-Made Trastuzumab (Herceptin) Inhibits HER2/Neu+ Cell Proliferation and Retards Tumor Growth

et al. 2011

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BackgroundPlant biotechnology provides a valuable contribution to global health, in part because it can decrease the cost of pharmaceutical products. Breast cancer can now be successfully treated by a humanized monoclonal antibody (mAb), trastuzumab (Herceptin). A course of treatment, however, is expensive and requires repeated administrations of the mAb. Here we used an Agrobacterium-mediated transient expression system to produce trastuzumab in plant cells.Methodology/Principal FindingsWe describe the cloning and expression of gene constructs in Nicotiana benthamiana plants using intron-optimized Tobacco mosaic virus- and Potato virus X-based vectors encoding, respectively, the heavy and light chains of trastuzumab. Full-size antibodies extracted and purified from plant tissues were tested for functionality and specificity by (i) binding to HER2/neu on the surface of a human mammary gland adenocarcinoma cell line, SK-BR-3, in fluorescence-activated cell sorting assay and (ii) testing the in vitro and in vivo inhibition of HER-2-expressing cancer cell proliferation. We show that plant-made trastuzumab (PMT) bound to the Her2/neu oncoprotein of SK-BR-3 cells and efficiently inhibited SK-BR-3 cell proliferation. Furthermore, mouse intraperitoneal PMT administration retarded the growth of xenografted tumors derived from human ovarian cancer SKOV3 Her2+ cells.Conclusions/SignificanceWe conclude that PMT is active in suppression of cell proliferation and tumor growth.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Plant-Made Trastuzumab (Herceptin) Inhibits HER2/Neu+ Cell Proliferation and Retards Tumor Growth

et al. 2011

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“…These antibodies carry only variable fragments and their relatively small size allows for easy genetic manipulation and molecular evolution to further improve their affinity [38–40]. In addition, fungus-specific scFvs, either alone or as a fusion with other molecules, have been shown to confer resistance to pathogens in transgenic plants [32–34]. …”

Section: Introductionmentioning

confidence: 99%

Generation of a Highly Reactive Chicken-Derived Single-Chain Variable Fragment against Fusarium verticillioides by Phage Display

Liu

et al. 2012

IJMS

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Fusarium verticillioides is the primary causal agent of Fusarium ear and kernel rot in maize, producing fumonisin mycotoxins that are toxic to humans and domestic animals. Rapid detection and monitoring of fumonisin-producing fungi are pivotally important for the prevention of mycotoxins from entering into food/feed products. Chicken-derived single-chain variable fragments (scFvs) against cell wall-bound proteins from F. verticillioides were isolated from an immunocompetent phage display library. Comparative phage enzyme-linked immunosorbant assays (ELISAs) and sequencing analyses identified four different scFv antibodies with high sensitivity. Soluble antibody ELISAs identified two highly sensitive scFv antibodies, FvCA3 and FvCA4, with the latter being slightly more sensitive. Three-dimensional modeling revealed that the FvCA4 may hold a better overall structure with CDRH3, CDRL1 and CDRL3 centered in the core region of antibody surface compared with that of other scFvs. Immunofluorescence labeling revealed that the binding of FvCA4 antibody was localized to the cell walls of conidiospores and hyphae of F. verticillioides, confirming the specificity of this antibody for a surface target. This scFv antibody was able to detect the fungal mycelium as low as 10−2 μg/mL and contaminating mycelium at a quantity of 10−2 mg/g maize. This is the first report that scFv antibodies derived from phage display have a wide application for rapid and accurate detection and monitoring of fumonisin-producing pathogens in agricultural samples.

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“…However, antibody-based resistance has been obtained even at low expression levels of cytosolic scFvs (Tavladoraki et al, 1993; Zimmermann et al, 1998). Some strategies have also been reported to overcome the instability of cytosolic expressed scFvs; for instance, inclusion of a C-terminal KDEL or other short polypeptides or construction of more stable and functional antibodies through phage display technology (De Jaeger et al, 1999; Jahromi et al, 2009; Prins et al, 2005; Schouten et al, 1996, 1997; Spiegel et al, 1999; Villani et al, 2005; Yajima et al, 2010). In our experience, both C and CC hairy roots targeting scFv to the cytosol with or without KDEL respectively, exhibited resistance against BNYVV (Table 1).…”

mentioning

confidence: 99%

Antibody-Mediated Resistance to Rhizomania Disease in Sugar Beet Hairy Roots

et al. 2019

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Expression of anti-sclerotinia scFv in transgenic Brassica napus enhances tolerance against stem rot

Cited by 23 publications

References 19 publications

Plant-Made Trastuzumab (Herceptin) Inhibits HER2/Neu+ Cell Proliferation and Retards Tumor Growth

Plant-Made Trastuzumab (Herceptin) Inhibits HER2/Neu+ Cell Proliferation and Retards Tumor Growth

Generation of a Highly Reactive Chicken-Derived Single-Chain Variable Fragment against Fusarium verticillioides by Phage Display

Antibody-Mediated Resistance to Rhizomania Disease in Sugar Beet Hairy Roots

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