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In Sik Chung - One of the best experts on this subject based on the ideXlab platform.
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Comparative Study on Trichoplusia ni Tn 5B1-4 Cells and Bombyx mori BmN Cells for Recombinant Endostatin Production
International journal of industrial entomology, 2020Co-Authors: Bong-hee Sohn, Pil-don Kang, In Sik ChungAbstract:The Recombinant plasmids harboring a heterologous gene coding mouse Endostatin were transfected and expressed stably in Trichoplusia ni Tn 5B1-4 cells and Bombyx mori BmN cells, respectively. Recombinant Endostatin expressed in the stably transformed Tn 5B1-4 and BmN cells was secreted into the medium. BmN cells are relatively lower in maximum cell growth and Recombinant Endostatin production than Tn 5B 1-4 cells. Recombinant Endostatin was also purified to homogeneity using a simple one-step affinity fractionation method. Purified Recombinant Endostatin inhibited endothelial cell proliferation in a dose-dependent manner. The concentration at half-maximum inhibition for Recombinant Endostatin was approximately 0.35 /ml.
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Functional expression of Recombinant canstatin in stably transformed Drosophila melanogaster S2 cells.
Protein Expression and Purification, 2006Co-Authors: Hwang-bo Jeon, Bong-hee Sohn, In Sik ChungAbstract:Recombinant tumstatin was expressed in stably transformed Drosophila melanogaster S2 cells and secreted into the medium with a molecular size of 29 kDa. Recombinant Endostatin was also purified to homogeneity using a simple one-step Ni2+ affinity fractionation. Purified Recombinant tumstatin inhibited endothelial cell proliferation in a dose-dependent manner. The concentration at half-maximum inhibition for Recombinant tumstatin was approx. 0.7 μg ml−1. A maximum production of 4.6 μg Recombinant tumstatin (107 cells)−1 was obtained in a T-flask culture of S2 cells, 7 d after induction with 0.5 mM CuSO4.
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Expression of Recombinant Endostatin in Agrobacterium-inoculated leaf disks of Nicotiana tabacum var. Xanthi
Biotechnology Letters, 2004Co-Authors: Seong Hyun Hong, Ho Yong Chung, Garry Sunter, David M. Bisaro, In Sik ChungAbstract:Recombinant Endostatin was transiently expressed in Agrobacterium -inoculated leaf disks of Nicotiana tabacum var. Xanthi with a molecular size of 23 kDa. Expression of Endostatin from a replicating vector based on tomato golden mosaic virus (TGMV) was 170% higher at the transcript level and double higher at the protein level than from a control vector of a non-replicating construct. Purified Recombinant Endostatin from tobacco leaf-disks has an anti-proliferative effect on bovine endothelial cells.
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Functional expression of Recombinant tumstatin in stably transformed Drosophila melanogaster S2 cells
Biotechnology Letters, 2003Co-Authors: Hee Kyoung Jeon, Kyung Hwa Chang, In Sik ChungAbstract:Recombinant tumstatin was expressed in stably transformed Drosophila melanogaster S2 cells and secreted into the medium with a molecular size of 29 kDa. Recombinant Endostatin was also purified to homogeneity using a simple one-step Ni^2+ affinity fractionation. Purified Recombinant tumstatin inhibited endothelial cell proliferation in a dose-dependent manner. The concentration at half-maximum inhibition for Recombinant tumstatin was approx. 0.7 μg ml^−1. A maximum production of 4.6 μg Recombinant tumstatin (10^7 cells)^−1 was obtained in a T-flask culture of S2 cells, 7 d after induction with 0.5 mM CuSO_4.
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Production of Recombinant Endostatin from stably transformed Trichoplusia ni BTI Tn 5B1-4 cells
Biotechnology Letters, 2001Co-Authors: Kyung Hwa Chang, Jong Hwa Park, In Sik ChungAbstract:The Recombinant plasmids harboring a heterologous gene coding mouse Endostatin were transfected and expressed stably in Trichoplusia ni BTI Tn 5B1-4 (Tn 5B1-4) cells. Recombinant Endostatin expressed in the stably transformed Tn 5B1-4 cells was secreted into the medium. Recombinant Endostatin was also purified to homogeneity using a simple one-step Ni^2+ affinity fractionation method. Purified Recombinant Endostatin inhibited endothelial cell proliferation in a dose-dependent manner. The concentration at half-maximum inhibition (ED_50) for Recombinant Endostatin was approximately 0.35 μg ml^−1. In a T-flask, the stably transformed Tn 5B1-4 cells produced 14.3 mg Recombinant Endostatin l^−1 at 6 days of cultivation.
Roy S. Herbst - One of the best experts on this subject based on the ideXlab platform.
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Study of the Media’s Potential Influence on Prospective Research Participants’ Understanding of and Motivations for Participation in a High-Profile Phase I Trial
Journal of Clinical Oncology, 2002Co-Authors: Rebecca D. Pentz, Anne L. Flamm, Jeremy Sugarman, Marlene Z. Cohen, G. Daniel Ayers, Roy S. Herbst, James L. AbbruzzeseAbstract:PURPOSE: To describe prospective participants’ initial source of information about, understanding of, and motivation to participate in a phase I clinical trial of the antiangiogenesis agent human Recombinant Endostatin. PATIENTS AND METHODS: We surveyed 100 of 130 persons referred to the Endostatin trial between October 1999 and November 2000 and analyzed media coverage of the agent from 1997 to 2000. RESULTS: Forty-seven percent of survey respondents first heard about the trial from media reports. Fifty-one percent of these subsequently contacted their physicians. Thirty-three percent of respondents correctly understood the purpose of the trial. Seventy-nine respondents were interviewed before they met trial investigators to discuss the trial. Of these, those who first heard about Endostatin from the media were five times more likely to understand correctly the trial’s purpose than those who first heard from other sources. Seventy-four percent (70 of 95) of respondents cited hope for personal benefit as ...
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development of biologic markers of response and assessment of antiangiogenic activity in a clinical trial of human Recombinant Endostatin
Journal of Clinical Oncology, 2002Co-Authors: Roy S. Herbst, Nizar A Mullani, Darren W Davis, Kenneth R Hess, David J Mcconkey, Chusilp Charnsangavej, Michael S Oreilly, Cheryl H Baker, Jennifer S Roach, Lee M EllisAbstract:PURPOSE: Angiogenesis is a target for the treatment of cancer and other diseases, and its complex biology suggests that establishing the appropriate dose and schedule for antiangiogenic treatment will require extensive study. We present the initial results of a dose-finding clinical trial of Recombinant human Endostatin (rh-Endo) that examined potential surrogates for response to antiangiogenic therapy. PATIENTS AND METHODS: Twenty-five patients were treated with escalating doses of rh-Endo. Positron emission tomography (PET) was used to assess tumor blood flow (with [15O]H2O) and metabolism (with [18F]fluorodeoxyglucose) before the start of therapy and then every 4 weeks. To directly assess the effects of rh-Endo on endothelial cells within the tumors, biopsy specimens of tumor tissue were obtained before therapy and again at 8 weeks and evaluated for endothelial cell and tumor cell apoptosis. RESULTS: Tumor blood flow and metabolism as measured by PET scans generally decreased with increasing doses of r...
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Targeted therapy using novel agents in the treatment of non-small-cell lung cancer.
Clinical Lung Cancer, 2002Co-Authors: Roy S. HerbstAbstract:Abstract Patients with advanced non—small-cell lung cancer (NSCLC) have a poor prognosis and high mortality. The therapeutic improvement caused by the new generation of cytotoxic agents seems to have reached a plateau. The main categories of targeted therapeutics applicable for NSCLC include receptor-targeted therapy, signal transduction or cell-cycle inhibition, angiogenesis inhibitors, gene therapy, and vaccines. Several major classes of agents directed at specific cellular mechanisms exist for the treatment of NSCLC. The anti—epidermal growth factor receptor (EGFR) group contains trastuzumab and IMC-C225, monoclonal antibodies against EGFRs that are overexpressed in many cancers. OSI-774 and ZD1839 are inhibitors of EGFR tyrosine kinase, a key enzyme of the signaling pathway. Farnesyl transferase inhibitors, such as SCH66336, and protein kinase C inhibitors, such as ISIS 3521, have also shown antitumor activity. Antiangiogenesis agents that have shown promise include TNP-470, Recombinant Endostatin, and angiostatin. Antibodies to vascular endothelial growth factor (VEGF) also seem to control tumor progression and may prolong survival. LY317615, an inhibitor of protein kinase Cβ, augmented the tumor growth delay produced by cytotoxic drugs. All of these agents are in different phases of clinical testing and have shown encouraging activity as single agents or in combination with chemotherapy drugs. These new agents are more target specific, less toxic, easier to administer, and may lead to enhanced safety and survival for patients with advanced NSCLC.
Jong Hwa Park - One of the best experts on this subject based on the ideXlab platform.
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Production of Recombinant Endostatin from stably transformed Trichoplusia ni BTI Tn 5B1-4 cells
Biotechnology Letters, 2001Co-Authors: Kyung Hwa Chang, Jong Hwa Park, In Sik ChungAbstract:The Recombinant plasmids harboring a heterologous gene coding mouse Endostatin were transfected and expressed stably in Trichoplusia ni BTI Tn 5B1-4 (Tn 5B1-4) cells. Recombinant Endostatin expressed in the stably transformed Tn 5B1-4 cells was secreted into the medium. Recombinant Endostatin was also purified to homogeneity using a simple one-step Ni^2+ affinity fractionation method. Purified Recombinant Endostatin inhibited endothelial cell proliferation in a dose-dependent manner. The concentration at half-maximum inhibition (ED_50) for Recombinant Endostatin was approximately 0.35 μg ml^−1. In a T-flask, the stably transformed Tn 5B1-4 cells produced 14.3 mg Recombinant Endostatin l^−1 at 6 days of cultivation.
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Optimal production and in vitro activity of recombinat Endostatin from stably transformed Drosophila melanogaster S2 cells
In Vitro Cellular & Developmental Biology - Animal, 2001Co-Authors: Jong Hwa Park, Kyung Hwa Chang, In Sik ChungAbstract:Recombinant plasmids containing a complementary deoxyribonucleic acid coding mouse Endostatin were transfected and stably expressed in Drosophila melanogaster Schneider 2 (S2) cells. Stably transformed polyclonal cell populations expressing Recombinant Endostatin were isolated after 4 wk of selection with hygromycin B. Recombinant Endostatin expressed in the stably transformed S2 cells under the influence of the Drosophila BiP protein signal sequence was secreted into the medium. Recombinant Endostatin was also purified to homogeneity using a simple one-step Ni^2+ affinity fractionation method. Purified Recombinant Endostatin inhibited endothelial cell proliferation in a dose-dependent manner. The concentration at maximum inhibition for Recombinant Endostatin was approximately 1.8 μg/ml. The stably transformed S2 cells produced 18 mg Recombinant Endostatin/L 7 d after induction with 5 μ M CdCl_2. Sodium butyrate supplementation (2.5 m M ) increased Recombinant Endostatin production by 17%. These findings demonstrate optimal production and in vitro activity of Recombinant Endostatin from stably transformation D. melanogaster S2 cells.
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Production of Recombinant Endostatin from stably transformed Drosophila melanogaster S2 cells
Biotechnology Letters, 1999Co-Authors: Jong Hwa Park, In Sik ChungAbstract:The Recombinant plasmids harboring a heterologous gene coding mouse Endostatin were transfected and expressed stably in Drosophila melanogaster S2 cells. Recombinant Endostatin expressed in the stably transformed S2 cells was secreted into the medium. Recombinant Endostatin was also purified to homogeneity using a simple one-step Ni^2+ affinity fractionation method. The purification yield was approximately 4 μg from the medium fraction of 8 ml cultures of stably transformed S2 cells. In a T-flask, the stably transformed S2 cells produced 24 mg Recombinant Endostatin/l at 7 days post-induction by 0.5 mM CuSO_4. In a high aspect rotating-wall vessel designed by NASA to simulate microgravity, the S2 cells produced up to 13 mg Recombinant Endostatin/l.
Kyung Hwa Chang - One of the best experts on this subject based on the ideXlab platform.
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Functional expression of Recombinant tumstatin in stably transformed Drosophila melanogaster S2 cells
Biotechnology Letters, 2003Co-Authors: Hee Kyoung Jeon, Kyung Hwa Chang, In Sik ChungAbstract:Recombinant tumstatin was expressed in stably transformed Drosophila melanogaster S2 cells and secreted into the medium with a molecular size of 29 kDa. Recombinant Endostatin was also purified to homogeneity using a simple one-step Ni^2+ affinity fractionation. Purified Recombinant tumstatin inhibited endothelial cell proliferation in a dose-dependent manner. The concentration at half-maximum inhibition for Recombinant tumstatin was approx. 0.7 μg ml^−1. A maximum production of 4.6 μg Recombinant tumstatin (10^7 cells)^−1 was obtained in a T-flask culture of S2 cells, 7 d after induction with 0.5 mM CuSO_4.
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Production of Recombinant Endostatin from stably transformed Trichoplusia ni BTI Tn 5B1-4 cells
Biotechnology Letters, 2001Co-Authors: Kyung Hwa Chang, Jong Hwa Park, In Sik ChungAbstract:The Recombinant plasmids harboring a heterologous gene coding mouse Endostatin were transfected and expressed stably in Trichoplusia ni BTI Tn 5B1-4 (Tn 5B1-4) cells. Recombinant Endostatin expressed in the stably transformed Tn 5B1-4 cells was secreted into the medium. Recombinant Endostatin was also purified to homogeneity using a simple one-step Ni^2+ affinity fractionation method. Purified Recombinant Endostatin inhibited endothelial cell proliferation in a dose-dependent manner. The concentration at half-maximum inhibition (ED_50) for Recombinant Endostatin was approximately 0.35 μg ml^−1. In a T-flask, the stably transformed Tn 5B1-4 cells produced 14.3 mg Recombinant Endostatin l^−1 at 6 days of cultivation.
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Optimal production and in vitro activity of recombinat Endostatin from stably transformed Drosophila melanogaster S2 cells
In Vitro Cellular & Developmental Biology - Animal, 2001Co-Authors: Jong Hwa Park, Kyung Hwa Chang, In Sik ChungAbstract:Recombinant plasmids containing a complementary deoxyribonucleic acid coding mouse Endostatin were transfected and stably expressed in Drosophila melanogaster Schneider 2 (S2) cells. Stably transformed polyclonal cell populations expressing Recombinant Endostatin were isolated after 4 wk of selection with hygromycin B. Recombinant Endostatin expressed in the stably transformed S2 cells under the influence of the Drosophila BiP protein signal sequence was secreted into the medium. Recombinant Endostatin was also purified to homogeneity using a simple one-step Ni^2+ affinity fractionation method. Purified Recombinant Endostatin inhibited endothelial cell proliferation in a dose-dependent manner. The concentration at maximum inhibition for Recombinant Endostatin was approximately 1.8 μg/ml. The stably transformed S2 cells produced 18 mg Recombinant Endostatin/L 7 d after induction with 5 μ M CdCl_2. Sodium butyrate supplementation (2.5 m M ) increased Recombinant Endostatin production by 17%. These findings demonstrate optimal production and in vitro activity of Recombinant Endostatin from stably transformation D. melanogaster S2 cells.
Ming-ying Zhen - One of the best experts on this subject based on the ideXlab platform.
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soluble multimer of Recombinant Endostatin expressed in e coli has anti angiogenesis activity
Biochemical and Biophysical Research Communications, 2006Co-Authors: Jian-fu Liang, Ming-ying ZhenAbstract:The bioactivity, refolding, and multimer formation of Endostatin, particularly of Recombinant Endostatin produced from bacteria, are proved challenging for clinical application. In order to determine the biological activity of Recombinant Endostatin multimer, first, we expressed Endostatin in Escherichia coli and purified it with ion-exchange chromatography. The purified active protein could elicit multimer formation spontaneously, but still has comparable activity. Aim to determine the anti-angiogenic activity of multimer Endostatin, by use of RP-HPLC, we then successfully separated Endostatin monomer and multimer for subjecting to anti-angiogenesis assay. The results from CAM (chorioallantoic membrane) inhibition assay showed that both monomer and multimer suppressed CAM vascularization significantly. At the dosage of 0.8 {mu}g, inhibition rates of multimeric and monomeric proteins were about 58% and 38%, respectively. Multimeric Endostatin exerted a higher activity than monomeric Endostatin (p 0.05), although both of them show a high inhibition effect in contrast to control. The results from HUVEC proliferation assay also showed similar effects at dosages of 0.6 and 1.6 {mu}g/ml, multimer exerted a higher activity on inhibition of HUVEC proliferation comparing with monomer (p
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Soluble multimer of Recombinant Endostatin expressed in E. coli has anti-angiogenesis activity.
Biochemical and Biophysical Research Communications, 2006Co-Authors: Jian-fu Liang, Ming-ying ZhenAbstract:The bioactivity, refolding, and multimer formation of Endostatin, particularly of Recombinant Endostatin produced from bacteria, are proved challenging for clinical application. In order to determine the biological activity of Recombinant Endostatin multimer, first, we expressed Endostatin in Escherichia coli and purified it with ion-exchange chromatography. The purified active protein could elicit multimer formation spontaneously, but still has comparable activity. Aim to determine the anti-angiogenic activity of multimer Endostatin, by use of RP-HPLC, we then successfully separated Endostatin monomer and multimer for subjecting to anti-angiogenesis assay. The results from CAM (chorioallantoic membrane) inhibition assay showed that both monomer and multimer suppressed CAM vascularization significantly. At the dosage of 0.8 {mu}g, inhibition rates of multimeric and monomeric proteins were about 58% and 38%, respectively. Multimeric Endostatin exerted a higher activity than monomeric Endostatin (p 0.05), although both of them show a high inhibition effect in contrast to control. The results from HUVEC proliferation assay also showed similar effects at dosages of 0.6 and 1.6 {mu}g/ml, multimer exerted a higher activity on inhibition of HUVEC proliferation comparing with monomer (p