Transdermal peptide conjugated to human connective tissue growth factor with enhanced cell proliferation and hyaluronic
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BIOTECHNOLOGICAL PRODUCTS AND PROCESS ENGINEERING
Transdermal peptide conjugated to human connective tissue growth factor with enhanced cell proliferation and hyaluronic acid synthesis activities produced by a silkworm silk gland bioreactor Yuancheng Wang 1,2 & Feng Wang 1,2,3 & Sheng Xu 1,2 & Riyuan Wang 1,2 & Chi Tian 1,2 & Yanting Ji 1,2 & Qianqian Yang 1,2 & Ping Zhao 1,2 & Qingyou Xia 1,2,3 Received: 30 April 2020 / Revised: 19 July 2020 / Accepted: 14 August 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
Abstract Human connective tissue growth factor (CTGF) is a secreted cysteine-rich peptide that stimulates cell proliferation, migration, and extracellular matrix production during tissue development, differentiation, angiogenesis, implantation, wound healing, and fibrosis processes, with broad application in the medical and cosmetic medical fields. However, the production of CTGF is currently limited by its low yield and purity in current bioreactors. In this study, two genetically modified silkworm strains were generated harboring artificially designed CTGF-8ht and pepCTGF-8ht genes, respectively, that contain an enhanced His-tag with eight histidine residues with or without a transdermal peptide (pep). Both recombinant CTGF-8ht and pepCTGF-8ht proteins were successfully expressed in the silkworm silk gland and cocoon, and could be easily extracted and purified from the cocoon by single-affinity immunoprecipitation column chromatography, achieving a purity of more than 95%. Moreover, compared with CTGF-8ht protein, pepCTGF-8ht protein exhibited better cell proliferation activity by activating the extracellular signalregulated kinase (ERK) pathway and enhanced hyaluronic acid synthesis activity by upregulating hyaluronan synthase 3 expression; moreover, the addition of pep significantly improved the transmembrane ability of CTGF-8ht protein. These results should help to promote the application prospects of CTGF and further guide the design and development of protein drugs from silkworm and other bioreactor systems. Key points & A silkworm bioreactor was optimized to produce connective tissue growth factor (CTGF) & The transgene contained an enhanced 8-His-tag and transmembrane peptide (pep) & Recombinant CTGF was easily purified with maintained or higher biological activity Keywords Human connective tissue growth factor . Transdermal peptide . Silkworm silk gland bioreactor . Genetic engineering
Introduction Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00253-020-10836-0) contains supplementary material, which is available to authorized users. * Qingyou Xia [email protected] 1
Biological Science Research Center, Southwest University, Chongqing 400715, People’s Republic of China
2
Chongqing Key Laboratory of Sericultural Science, Southwest University, Chongqing 400715, People’s Republic of China
3
Chongqing Engineering and Technology Research Center for Novel Silk Materials, Southwest University, Chongqing 400715, People’s Republic of C
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