The Experts below are selected from a list of 7449 Experts worldwide ranked by ideXlab platform
Yun-gen Miao - One of the best experts on this subject based on the ideXlab platform.
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Probability to produce animal vaccines in insect Baculovirus Expression System
African Journal of Biotechnology, 2011Co-Authors: Shuang Liang, Peng Zhang, Fang Zhou, Yun-gen MiaoAbstract:The insect Baculovirus Expression System is a valuable tool for the production of vaccine. Many subunit vaccines have been expressed in this System. The first vaccine produced in insect cells for animal use is now in the market. In this study, we reviewed recent progress of animal’s vaccine production for different Expression levels and Baculovirus genome stability, characteristic features of Baculovirus Expression vector System (BVES), virus link particles, Baculovirus Expression in mammalian cell and methodology of produce subunit vaccines. This review showed that BVES is a fantastic tool for vaccine development and it has wonderful feature for future animal vaccine development.
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cloning and Expression of a cellulase gene in the silkworm bombyx mori by improved bac to bac bmnpv Baculovirus Expression System
Molecular Biology Reports, 2010Co-Authors: Xinghua Li, Fang Zhou, Dan Wang, Huajun Yang, Roy Bhaskar, Jiabiao Hu, Yun-gen MiaoAbstract:Cellulases catalyze the hydrolysis of cellulose which are mainly three types: endoglucanases, cellobiohydrolases and β-glucosidases. It can be used in converting cellulosic biomass to glucose that can be used in different applications such as production of fuel ethanol, animal feed, waste water treatment and in brewing industry. In this paper, we cloned a 1380-bp endoglucanase I (EG I) gene from mycelium of filamentous fungus Trichoderma viride strain AS 3.3711 using PCR-based exon splicing methods, and expressed the recombinant EG I mature peptide protein in both silkworm BmN cell line and silkworm larvae with a newly established Bac-to-Bac/BmNPV mutant Baculovirus Expression System, which lacks the virus-encoded chitinase (chiA) and cathepsin (v-cath) genes of Bombyx mori nucleopolyhedrovirus (BmNPV). An around 49-kDa protein was visualized after mBacmid/BmNPV/EG I infection, and the maximum Expression in silkworm larvae was at 84 h post-infection. The ANOVA showed that the enzymes from recombinant Baculoviruses infected silkworms exhibited significant maximum enzyme activity at the environmental condition of pH 7.0 and temperature 50°C. It was stable at pH range from 5.0 to 10.0 and at temperature range from 50 to 60°C, and increased 24.71 and 22.84% compared with that from wild Baculoviruses infected silkworms and normal silkworms, respectively. The availability of large quantities of EG I that the silkworm provides maybe greatly facilitate the future research and the potential application in industries.
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cloning and Expression of manganese superoxide dismutase of the silkworm bombyx mori by bac to bac bmnpv Baculovirus Expression System
Applied Microbiology and Biotechnology, 2006Co-Authors: Yun-gen Miao, Xiaofeng Wu, Aichun Zhao, Xinghua Li, Masao NakagakiAbstract:Superoxide dismutase (SODs) are metalloenzymes that catalyze the dismutation of the superoxide anion to molecular oxygen and hydrogen peroxide and, thus, form a crucial part of the cellular antioxidant defense mechanism. In this paper, we used the total fat body RNA of silkworm, Bombyx mori L. to clone and sequence a 648-bp Mn-SOD cDNA fragment through RT-PCR. Furthermore, a newly established Bac-to-Bac/BmNPV Baculovirus Expression System was used to overexpress the recombinant Mn-SOD enzyme in silkworm larvae. The hemolymph was collected from the infected larvae 96 h post-infection and subjected to a 12 % SDS-PAGE and Western blotting. A 18.0-kDa protein was visualized after rBacmid/BmNPV/SOD infection. The SOD enzyme activity was determined with a tetrazolium salt for detection of superoxide radicals generated by xanthine and xanthine oxidase and its peak appeared in 96 h post-infection with 2.7 times of the control larvae. The availability of large quantities of SOD that the silkworm provides should greatly facilitate the future research and testing of this protein for potential application in medicine.
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Expression of spider flagelliform silk protein in bombyx mori cell line by a novel bac to bac bmnpv Baculovirus Expression System
Applied Microbiology and Biotechnology, 2006Co-Authors: Yun-gen Miao, Yuansong Zhang, Koichi Nakagaki, Tianfu Zhao, Aichun Zhao, Yan Meng, Masao Nakagaki, Enoch Y Park, Katsumi MaenakaAbstract:Bombyx mori nuclear polyhedrosis virus (BmNPV) Baculovirus Expression System (BES) has a lot of advantages such as high Expression efficiency, convenience, and low feeding cost. In this report, we used a recently developed BmNPV bacmid, which could infect both B. mori cell lines and silkworm larvae. The results showed it takes only 7 to 10 days to generate recombinant Baculovirus and permit the rapid isolation from small-scale cultures and then use it to transfect B. mori cell lines, compared to traditional homologous recombination method, which needs at least 40 days for multiple rounds of purification and amplification of viruses. Using this BES, we expressed a recombinant spider flagelliform protein in BmN cell line, which was around 37 kDa in sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and western blot analysis. The BmNPV bacmid System using silkworm would be very attractive for Expression of target proteins.
Kouhei Tsumoto - One of the best experts on this subject based on the ideXlab platform.
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Characterization of glycoengineered anti-HER2 monoclonal antibodies produced by using a silkworm–Baculovirus Expression System
Journal of Biochemistry, 2018Co-Authors: Yuriko Egashira, Satoru Nagatoishi, Masato Kiyoshi, Akiko Ishii-watabe, Kouhei TsumotoAbstract:Silkworm-Baculovirus Expression Systems are efficient means for the production of recombinant proteins that provide high Expression levels and post-translational modifications. Here, we characterized the stability, glycosylation pattern and antibody-dependent cell-mediated cytotoxicity activity of anti-HER2 monoclonal antibodies containing native or glycoengineered mammalian-like N-glycans that were produced by using a silkworm-Baculovirus Expression System. Compared with a monoclonal antibody produced by using a Chinese hamster ovary cell Expression System, the glycoengineered monoclonal antibody had comparable thermal stability and a higher antibody-dependent cell-mediated cytotoxicity activity. These results suggest that silkworm-Baculovirus Expression Systems are next-generation Expression Systems potentially useful for the cost-effective production of therapeutic antibodies.
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Characterization of glycoengineered anti-HER2 monoclonal antibodies produced by using a silkworm-Baculovirus Expression System
Journal of Biochemistry, 2018Co-Authors: Yuriko Egashira, Satoru Nagatoishi, Masato Kiyoshi, Akiko Ishii-watabe, Kouhei TsumotoAbstract:© The Author(s) 2018. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved. Silkworm-Baculovirus Expression Systems are efficient means for the production of recombinant proteins that provide high Expression levels and post-translational modifications. Here, we characterized the stability, glycosylation pattern and antibody-dependent cell-mediated cytotoxicity activity of anti-HER2 monoclonal antibodies containing native or glycoengineered mammalian-like N-glycans that were produced by using a silkworm-Baculovirus Expression System. Compared with a monoclonal antibody produced by using a Chinese hamster ovary cell Expression System, the glycoengineered monoclonal antibody had comparable thermal stability and a higher antibody-dependent cell-mediated cytotoxicity activity. These results suggest that silkworm-Baculovirus Expression Systems are next-generation Expression Systems potentially useful for the cost-effective production of therapeutic antibodies.
Masao Nakagaki - One of the best experts on this subject based on the ideXlab platform.
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cloning and Expression of manganese superoxide dismutase of the silkworm bombyx mori by bac to bac bmnpv Baculovirus Expression System
Applied Microbiology and Biotechnology, 2006Co-Authors: Yun-gen Miao, Xiaofeng Wu, Aichun Zhao, Xinghua Li, Masao NakagakiAbstract:Superoxide dismutase (SODs) are metalloenzymes that catalyze the dismutation of the superoxide anion to molecular oxygen and hydrogen peroxide and, thus, form a crucial part of the cellular antioxidant defense mechanism. In this paper, we used the total fat body RNA of silkworm, Bombyx mori L. to clone and sequence a 648-bp Mn-SOD cDNA fragment through RT-PCR. Furthermore, a newly established Bac-to-Bac/BmNPV Baculovirus Expression System was used to overexpress the recombinant Mn-SOD enzyme in silkworm larvae. The hemolymph was collected from the infected larvae 96 h post-infection and subjected to a 12 % SDS-PAGE and Western blotting. A 18.0-kDa protein was visualized after rBacmid/BmNPV/SOD infection. The SOD enzyme activity was determined with a tetrazolium salt for detection of superoxide radicals generated by xanthine and xanthine oxidase and its peak appeared in 96 h post-infection with 2.7 times of the control larvae. The availability of large quantities of SOD that the silkworm provides should greatly facilitate the future research and testing of this protein for potential application in medicine.
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Expression of spider flagelliform silk protein in bombyx mori cell line by a novel bac to bac bmnpv Baculovirus Expression System
Applied Microbiology and Biotechnology, 2006Co-Authors: Yun-gen Miao, Yuansong Zhang, Koichi Nakagaki, Tianfu Zhao, Aichun Zhao, Yan Meng, Masao Nakagaki, Enoch Y Park, Katsumi MaenakaAbstract:Bombyx mori nuclear polyhedrosis virus (BmNPV) Baculovirus Expression System (BES) has a lot of advantages such as high Expression efficiency, convenience, and low feeding cost. In this report, we used a recently developed BmNPV bacmid, which could infect both B. mori cell lines and silkworm larvae. The results showed it takes only 7 to 10 days to generate recombinant Baculovirus and permit the rapid isolation from small-scale cultures and then use it to transfect B. mori cell lines, compared to traditional homologous recombination method, which needs at least 40 days for multiple rounds of purification and amplification of viruses. Using this BES, we expressed a recombinant spider flagelliform protein in BmN cell line, which was around 37 kDa in sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and western blot analysis. The BmNPV bacmid System using silkworm would be very attractive for Expression of target proteins.
Xinghua Li - One of the best experts on this subject based on the ideXlab platform.
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cloning and Expression of a cellulase gene in the silkworm bombyx mori by improved bac to bac bmnpv Baculovirus Expression System
Molecular Biology Reports, 2010Co-Authors: Xinghua Li, Fang Zhou, Dan Wang, Huajun Yang, Roy Bhaskar, Jiabiao Hu, Yun-gen MiaoAbstract:Cellulases catalyze the hydrolysis of cellulose which are mainly three types: endoglucanases, cellobiohydrolases and β-glucosidases. It can be used in converting cellulosic biomass to glucose that can be used in different applications such as production of fuel ethanol, animal feed, waste water treatment and in brewing industry. In this paper, we cloned a 1380-bp endoglucanase I (EG I) gene from mycelium of filamentous fungus Trichoderma viride strain AS 3.3711 using PCR-based exon splicing methods, and expressed the recombinant EG I mature peptide protein in both silkworm BmN cell line and silkworm larvae with a newly established Bac-to-Bac/BmNPV mutant Baculovirus Expression System, which lacks the virus-encoded chitinase (chiA) and cathepsin (v-cath) genes of Bombyx mori nucleopolyhedrovirus (BmNPV). An around 49-kDa protein was visualized after mBacmid/BmNPV/EG I infection, and the maximum Expression in silkworm larvae was at 84 h post-infection. The ANOVA showed that the enzymes from recombinant Baculoviruses infected silkworms exhibited significant maximum enzyme activity at the environmental condition of pH 7.0 and temperature 50°C. It was stable at pH range from 5.0 to 10.0 and at temperature range from 50 to 60°C, and increased 24.71 and 22.84% compared with that from wild Baculoviruses infected silkworms and normal silkworms, respectively. The availability of large quantities of EG I that the silkworm provides maybe greatly facilitate the future research and the potential application in industries.
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cloning and Expression of manganese superoxide dismutase of the silkworm bombyx mori by bac to bac bmnpv Baculovirus Expression System
Applied Microbiology and Biotechnology, 2006Co-Authors: Yun-gen Miao, Xiaofeng Wu, Aichun Zhao, Xinghua Li, Masao NakagakiAbstract:Superoxide dismutase (SODs) are metalloenzymes that catalyze the dismutation of the superoxide anion to molecular oxygen and hydrogen peroxide and, thus, form a crucial part of the cellular antioxidant defense mechanism. In this paper, we used the total fat body RNA of silkworm, Bombyx mori L. to clone and sequence a 648-bp Mn-SOD cDNA fragment through RT-PCR. Furthermore, a newly established Bac-to-Bac/BmNPV Baculovirus Expression System was used to overexpress the recombinant Mn-SOD enzyme in silkworm larvae. The hemolymph was collected from the infected larvae 96 h post-infection and subjected to a 12 % SDS-PAGE and Western blotting. A 18.0-kDa protein was visualized after rBacmid/BmNPV/SOD infection. The SOD enzyme activity was determined with a tetrazolium salt for detection of superoxide radicals generated by xanthine and xanthine oxidase and its peak appeared in 96 h post-infection with 2.7 times of the control larvae. The availability of large quantities of SOD that the silkworm provides should greatly facilitate the future research and testing of this protein for potential application in medicine.
Yuriko Egashira - One of the best experts on this subject based on the ideXlab platform.
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Characterization of glycoengineered anti-HER2 monoclonal antibodies produced by using a silkworm–Baculovirus Expression System
Journal of Biochemistry, 2018Co-Authors: Yuriko Egashira, Satoru Nagatoishi, Masato Kiyoshi, Akiko Ishii-watabe, Kouhei TsumotoAbstract:Silkworm-Baculovirus Expression Systems are efficient means for the production of recombinant proteins that provide high Expression levels and post-translational modifications. Here, we characterized the stability, glycosylation pattern and antibody-dependent cell-mediated cytotoxicity activity of anti-HER2 monoclonal antibodies containing native or glycoengineered mammalian-like N-glycans that were produced by using a silkworm-Baculovirus Expression System. Compared with a monoclonal antibody produced by using a Chinese hamster ovary cell Expression System, the glycoengineered monoclonal antibody had comparable thermal stability and a higher antibody-dependent cell-mediated cytotoxicity activity. These results suggest that silkworm-Baculovirus Expression Systems are next-generation Expression Systems potentially useful for the cost-effective production of therapeutic antibodies.
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Characterization of glycoengineered anti-HER2 monoclonal antibodies produced by using a silkworm-Baculovirus Expression System
Journal of Biochemistry, 2018Co-Authors: Yuriko Egashira, Satoru Nagatoishi, Masato Kiyoshi, Akiko Ishii-watabe, Kouhei TsumotoAbstract:© The Author(s) 2018. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved. Silkworm-Baculovirus Expression Systems are efficient means for the production of recombinant proteins that provide high Expression levels and post-translational modifications. Here, we characterized the stability, glycosylation pattern and antibody-dependent cell-mediated cytotoxicity activity of anti-HER2 monoclonal antibodies containing native or glycoengineered mammalian-like N-glycans that were produced by using a silkworm-Baculovirus Expression System. Compared with a monoclonal antibody produced by using a Chinese hamster ovary cell Expression System, the glycoengineered monoclonal antibody had comparable thermal stability and a higher antibody-dependent cell-mediated cytotoxicity activity. These results suggest that silkworm-Baculovirus Expression Systems are next-generation Expression Systems potentially useful for the cost-effective production of therapeutic antibodies.
