增强了人类角质细胞生长因子的核细胞表达,净化和生物活动
Thi Luong Vu1, Thi Kieu Oanh Nguyen1, Jung-A Song1
1Department of Physiology, University of Ulsan College of Medicine, Asan Medical Center, Seoul 05505, South Korea.
Journal of biotechnology
|March 29, 2024
概括
研究人员使用大肠杆菌中的蛋白质标签改善了复合皮细胞生长因子 (KGF) 的产生. 增强的KGF蛋白显示出显著的细胞增殖和伤口愈合能力,具有治疗潜力.
科学领域:
- 生物技术是生物技术.
- 分子生物学分子生物学
- 细胞生物学 细胞生物学
背景情况:
- 角质细胞生长因子 (KGF),也称为纤维细胞生长因子7 (FGF7),对细胞发育和细胞调节至关重要.
- 重组KGF生产的挑战包括在大肠杆菌中低表达和聚合.
研究的目的:
- 为了增强复合KGF的表达和溶解性.
- 评估净化KGF的生物活性.
主要方法:
- 工程化KGF与N端蛋白标签:PDIb'a',MBP,以及他的.
- 选择H-PDIb'a'-KGF用于大规模生产和净化.
- 使用液体染色学-并联质谱法 (LC-MS/MS) 确认纯度和同一性.
- 通过细胞增殖试验 (MCF-7细胞) 和伤口愈合试验 (HaCaT细胞) 评估生物活性.
主要成果:
- H-PDIb'a'-KGF融合蛋白显示出卓越的稳定性和表达.
- 纯化的KGF显著刺激了MCF-7细胞的增殖 (EC50 = 6.4 ± 0.5 pM).
- 单独的PDIb'a'也对MCF-7细胞产生了刺激作用.
- 重组KGF剂量依赖增强了HaCaT角质细胞伤口愈合.
结论:
- 开发了一种生产可溶和活性重组KGF的高效策略.
- 证明了纯化KGF在促进细胞增殖和伤口愈合方面的生物学有效性.
- 这些发现支持重组KGF的潜在治疗应用.
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