Effect of nitrogen deficiency on recombinant protein production and dimerization and growth in transgenic plants

Kang, Yangjoo; Shin, Yong Kyoo; Park, Sang-Won; Ko, Kisung

doi:10.1007/s13580-016-0045-5

Cited by 8 publications

(9 citation statements)

References 33 publications

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“…The recombinant plant expression vector was transferred into Agrobacterium tumefaciens strain LBA4404 by electroporation. Transgenic tobacco (Nicotiana tabacum Xanthi) plants were generated by Agrobacterium-mediated transformation [33]. The in vitro cultivated tobacco leaves were cut in two or three pieces and incubated with Murashige and Skoog (MS) medium for 1 min with gentle shaking.…”

Section: Plant Transformationmentioning

confidence: 99%

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Expression and In Vitro Function of Anti-Breast Cancer Llama-Based Single Domain Antibody VHH Expressed in Tobacco Plants

Park

Lee

Kim

et al. 2020

IJMS

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Overexpression of human epidermal growth factor receptor type 2 (HER2) is considered as a prognostic factor of breast cancer, which is positively associated with recurrence when cancer metastasizes to the lymph nodes. Here, we expressed the single variable domain on a heavy chain (VHH) form of anti-HER2 camelid single domain antibody in tobacco plants and compared its in vitro anticancer activities with the anti-HER2 full size antibody. The gene expression cassette containing anti-HER2 camelid single domain antibody VHH fused to human IgG Fc region with KDEL endoplasmic reticulum (ER) (VHH-FcK) was transferred into the tobacco plant via the Agrobacterium-mediated transformation. The transformants were screened with polymerase chain reaction and Western blot analyses. Enzyme-linked immunosorbent assay (ELISA) confirmed the binding of the purified anti-HER2 VHH-FcK to the HER2-positive breast cancer cell line, SK-BR-3. Migration assay results confirmed anticancer activity of the plant-derived anticancer camelid single chain antibody. Taken together, we confirmed the possibility of using anti-HER2 VHH-FcK as a therapeutic anticancer agent, which can be expressed and assembled and purified from a plant expression system as an alternative antibody production system.

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Section: Plant Transformationmentioning

confidence: 99%

“…The transgenic plants were transferred to a greenhouse to obtain whole plants. Transgenic and nontransgenic plants were cultivated in greenhouse under controlled conditions [33].…”

Section: Plant Transformationmentioning

confidence: 99%

Expression and In Vitro Function of Anti-Breast Cancer Llama-Based Single Domain Antibody VHH Expressed in Tobacco Plants

Park

Lee

Kim

et al. 2020

IJMS

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“…Agrobacterium -mediated transformation is the main tool to transfer the transfer DNA (T-DNA) region, which contains the gene expression cassette consisting of promoter, recombinant vaccine protein gene of interest, and terminator, into the plant genomic DNA. This technique has allowed the stable transformation of many plant species to express recombinant anti-cancer vaccine proteins ( Brodzik et al ., 2008 ; Lu et al ., 2012 ; Lim et al ., 2015 ; Kang et al ., 2016 ; Kim et al ., 2016 ).…”

Section: Plant Expression Systems For Vaccine Productionmentioning

confidence: 99%

“…Blue light (440 nm) and far red light (660 nm) are selectively applied as an energy source for photosynthesis to enable plant growth ( Singh et al ., 2015 ). Environmental factors such as temperature, soil nutrition, salinity, and drought stress all affect glycosylation, as well as protein qualities and quantities ( Jamal et al ., 2009 ; Kang et al ., 2016 ). Glycosylation patterns in the anti-cancer vaccine GA733-FcK in tobacco vary at the developmental stages ( Lim et al ., 2015 ).…”

Section: Effect Of Developmental and Environmental Factors On Vaccinementioning

confidence: 99%

Production of Recombinant Anti-Cancer Vaccines in Plants

Lee

2017

Biomolecules & Therapeutics

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Plant expression systems have been developed to produce anti-cancer vaccines. Plants have several advantages as bioreactors for the production of subunit vaccines: they are considered safe, and may be used to produce recombinant proteins at low production cost. However, several technical issues hinder large-scale production of anti-cancer vaccines in plants. The present review covers design strategies to enhance the immunogenicity and therapeutic potency of anti-cancer vaccines, methods to increase vaccine-expressing plant biomass, and challenges facing the production of anti-cancer vaccines in plants. Specifically, the issues such as low expression levels and plant-specific glycosylation are described, along with their potential solutions.

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“…One option to ensure cost-effective production of antibodies is the use of a plant expression system without pathogenic animal contaminants, high-priced manufacturing process, and expensive culture media [ 22 , 23 , 24 , 25 , 26 ]. The advantages of plant expression systems include inexpensive inputs (sunlight, water, and nutrients), the ease of large-scale production by increasing the area located in the seed, and low technology harvest compared with mammalian expression systems [ 22 , 27 , 28 , 29 ]. Moreover, plant expression systems are suitable for post-translational modifications of proteins, including the N -glycosylation required for biological activity, similar to mammalian expression systems [ 25 , 28 , 30 , 31 ].…”

Section: Introductionmentioning

confidence: 99%

Expression of a Large Single-Chain 13F6 Antibody with Binding Activity against Ebola Virus-Like Particles in a Plant System

Lim

Kim

2020

IJMS

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Pathogenic animal and human viruses present a growing and persistent threat to humans worldwide. Ebola virus (EBOV) causes zoonosis in humans. Here, two structurally different anti-Ebola 13F6 antibodies, recognizing the heavily glycosylated mucin-like domain (MLD) of the glycoprotein (GP), were expressed in transgenic Nicotiana tabacum plants and designed as inexpensive and effective diagnostic antibodies against Ebola virus disease (EVD). The first was anti-EBOV 13F6 full size antibody with heavy chain (HC) and light chain (LC) (monoclonal antibody, mAb 13F6-FULL), while the second was a large single-chain (LSC) antibody (mAb 13F6-LSC). mAb 13F6-LSC was constructed by linking the 13F6 LC variable region (VL) with the HC of mAb 13F6-FULL using a peptide linker and extended to the C-terminus using the endoplasmic reticulum (ER) retention motif KDEL. Agrobacterium-mediated plant transformation was employed to express the antibodies in N. tabacum. PCR, RT-PCR, and immunoblot analyses confirmed the gene insertion, transcription, and protein expression of these antibodies, respectively. The antibodies tagged with the KDEL motif displayed high-mannose type N-glycan structures and efficient binding to EBOV-like particles (VLPs). Thus, various forms of anti-EBOV plant-derived mAbs 13F6-FULL and LSC with efficient binding affinity to EBOV VLP can be produced in the plant system.

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Effect of nitrogen deficiency on recombinant protein production and dimerization and growth in transgenic plants

Cited by 8 publications

References 33 publications

Expression and In Vitro Function of Anti-Breast Cancer Llama-Based Single Domain Antibody VHH Expressed in Tobacco Plants

Expression and In Vitro Function of Anti-Breast Cancer Llama-Based Single Domain Antibody VHH Expressed in Tobacco Plants

Production of Recombinant Anti-Cancer Vaccines in Plants

Expression of a Large Single-Chain 13F6 Antibody with Binding Activity against Ebola Virus-Like Particles in a Plant System

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