Bulaon Christine Joy I, Shanmugaraj Balamurugan, Oo Yamin, Rattanapisit Kaewta, Chuanasa Taksina, Chaotham Chatchai, Phoolcharoen Waranyoo
Department of Pharmacognosy and Pharmaceutical Botany, Faculty of Pharmaceutical Sciences, Chulalongkorn University, Bangkok, Thailand.
Research Unit for Plant-produced Pharmaceuticals, Chulalongkorn University, Bangkok, Thailand.
Biotechnol Rep (Amst). 2020 Aug 15;27:e00514. doi: 10.1016/j.btre.2020.e00514. eCollection 2020 Sep.
Human vascular endothelial growth factor (VEGF) is a potent pro-angiogenic growth factor essential for wound healing. Due to its potential applications, many expression strategies have been developed to produce high levels of VEGF. Here, we have optimized the expression conditions for the production of recombinant VEGF in by using a geminiviral vector. Four different expression constructs that differ by the location of a C- or N-terminal histidine tag and SEKDEL sequence were developed and utilized for plant transient expression. The recombinant VEGF was further purified by using affinity chromatography and confirmed by SDS-PAGE and Western blotting probed with anti-VEGF antibody. Furthermore, our results showed that the recombinant VEGF in all tested concentrations did not exhibit any cytotoxic effect on HaCaT cells and induced cell migration . These findings show that the plant-produced VEGF has the potential to be used in regenerative medicine and cosmetic industry.
人血管内皮生长因子(VEGF)是伤口愈合所必需的一种强效促血管生成生长因子。由于其潜在应用价值,人们已开发出多种表达策略来大量生产VEGF。在此,我们通过使用双生病毒载体优化了在[具体内容缺失]中生产重组VEGF的表达条件。开发了四种不同的表达构建体,它们因C端或N端组氨酸标签以及SEKDEL序列的位置不同而有所差异,并用于植物瞬时表达。重组VEGF通过亲和层析进一步纯化,并用抗VEGF抗体进行SDS-PAGE和Western印迹分析加以确认。此外,我们的结果表明,所有测试浓度的重组VEGF对HaCaT细胞均未表现出任何细胞毒性作用,并能诱导细胞迁移。这些发现表明,植物产生的VEGF有潜力用于再生医学和化妆品行业。