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Transgenic Silkworm-Based Silk Gland Bioreactor for Large Scale Production of Bioactive Human Platelet-Derived Growth Factor (PDGF-BB) in Silk Cocoons

Human platelet derived growth factor (PDGF) is a major therapeutic protein with great demand in the clinical setting; however, its rate of supply is far from meeting needs. Here, we provide an effective strategy to produce PDGF-BB in large quantities using a transgenic silkworm. The codon-optimized...

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Autores principales: Chen, Wenjing, Wang, Feng, Tian, Chi, Wang, Yuancheng, Xu, Sheng, Wang, Riyuan, Hou, Kai, Zhao, Ping, Yu, Ling, Lu, Zhisong, Xia, Qingyou
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6164493/
https://www.ncbi.nlm.nih.gov/pubmed/30150526
http://dx.doi.org/10.3390/ijms19092533
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author Chen, Wenjing
Wang, Feng
Tian, Chi
Wang, Yuancheng
Xu, Sheng
Wang, Riyuan
Hou, Kai
Zhao, Ping
Yu, Ling
Lu, Zhisong
Xia, Qingyou
author_facet Chen, Wenjing
Wang, Feng
Tian, Chi
Wang, Yuancheng
Xu, Sheng
Wang, Riyuan
Hou, Kai
Zhao, Ping
Yu, Ling
Lu, Zhisong
Xia, Qingyou
author_sort Chen, Wenjing
collection PubMed
description Human platelet derived growth factor (PDGF) is a major therapeutic protein with great demand in the clinical setting; however, its rate of supply is far from meeting needs. Here, we provide an effective strategy to produce PDGF-BB in large quantities using a transgenic silkworm. The codon-optimized PDGF-B gene regulated by the highly efficient sericin-1 expression system was integrated into the genome of a silkworm. The high transcriptional expression of the PDGF-BB gene in the transgenic silkworm competitively inhibited the transcription expression of the endogenous sericin-1 gene which caused a significant 37.5% decline. The PDGF-BB synthesized in the middle silk gland (MSG) of transgenic silkworms could form a homodimer through intermolecular disulfide bonds, which is then secreted into sericin lumen and finally, distributed in the sericin layer of the cocoon. In this study, a protein quantity of approximately 0.33 mg/g was found in the cocoon. Following a purification process, approximately 150.7 μg of recombinant PDGF-BB with a purity of 82% was purified from 1 g of cocoons. Furthermore, the bioactivity assays showed that the purified recombinant PDGF-BB was able to promote the growth, proliferation and migration of NIH/3T3 cells significantly. These results suggest that the silk gland bioreactor can produce active recombinant PDGF-BB as an efficient mitogen and wound healing agent.
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spelling pubmed-61644932018-10-10 Transgenic Silkworm-Based Silk Gland Bioreactor for Large Scale Production of Bioactive Human Platelet-Derived Growth Factor (PDGF-BB) in Silk Cocoons Chen, Wenjing Wang, Feng Tian, Chi Wang, Yuancheng Xu, Sheng Wang, Riyuan Hou, Kai Zhao, Ping Yu, Ling Lu, Zhisong Xia, Qingyou Int J Mol Sci Article Human platelet derived growth factor (PDGF) is a major therapeutic protein with great demand in the clinical setting; however, its rate of supply is far from meeting needs. Here, we provide an effective strategy to produce PDGF-BB in large quantities using a transgenic silkworm. The codon-optimized PDGF-B gene regulated by the highly efficient sericin-1 expression system was integrated into the genome of a silkworm. The high transcriptional expression of the PDGF-BB gene in the transgenic silkworm competitively inhibited the transcription expression of the endogenous sericin-1 gene which caused a significant 37.5% decline. The PDGF-BB synthesized in the middle silk gland (MSG) of transgenic silkworms could form a homodimer through intermolecular disulfide bonds, which is then secreted into sericin lumen and finally, distributed in the sericin layer of the cocoon. In this study, a protein quantity of approximately 0.33 mg/g was found in the cocoon. Following a purification process, approximately 150.7 μg of recombinant PDGF-BB with a purity of 82% was purified from 1 g of cocoons. Furthermore, the bioactivity assays showed that the purified recombinant PDGF-BB was able to promote the growth, proliferation and migration of NIH/3T3 cells significantly. These results suggest that the silk gland bioreactor can produce active recombinant PDGF-BB as an efficient mitogen and wound healing agent. MDPI 2018-08-27 /pmc/articles/PMC6164493/ /pubmed/30150526 http://dx.doi.org/10.3390/ijms19092533 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Chen, Wenjing
Wang, Feng
Tian, Chi
Wang, Yuancheng
Xu, Sheng
Wang, Riyuan
Hou, Kai
Zhao, Ping
Yu, Ling
Lu, Zhisong
Xia, Qingyou
Transgenic Silkworm-Based Silk Gland Bioreactor for Large Scale Production of Bioactive Human Platelet-Derived Growth Factor (PDGF-BB) in Silk Cocoons
title Transgenic Silkworm-Based Silk Gland Bioreactor for Large Scale Production of Bioactive Human Platelet-Derived Growth Factor (PDGF-BB) in Silk Cocoons
title_full Transgenic Silkworm-Based Silk Gland Bioreactor for Large Scale Production of Bioactive Human Platelet-Derived Growth Factor (PDGF-BB) in Silk Cocoons
title_fullStr Transgenic Silkworm-Based Silk Gland Bioreactor for Large Scale Production of Bioactive Human Platelet-Derived Growth Factor (PDGF-BB) in Silk Cocoons
title_full_unstemmed Transgenic Silkworm-Based Silk Gland Bioreactor for Large Scale Production of Bioactive Human Platelet-Derived Growth Factor (PDGF-BB) in Silk Cocoons
title_short Transgenic Silkworm-Based Silk Gland Bioreactor for Large Scale Production of Bioactive Human Platelet-Derived Growth Factor (PDGF-BB) in Silk Cocoons
title_sort transgenic silkworm-based silk gland bioreactor for large scale production of bioactive human platelet-derived growth factor (pdgf-bb) in silk cocoons
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6164493/
https://www.ncbi.nlm.nih.gov/pubmed/30150526
http://dx.doi.org/10.3390/ijms19092533
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