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一种量化方法来区分细胞质与内体捕获的细胞透
Françoise Illien1, Zoltán Bánóczi2, Sandrine Sagan1
1Sorbonne Université, École normale supérieure, PSL University, CNRS, Laboratoire des Biomolécules, LBM, 75005, Paris, France.
Chembiochem : a European journal of chemical biology
|April 8, 2024
概括
我们开发了一种新的测试方法,以区分通过内细胞结合 (endocytosis) 和直接转移 (direct translocation) 的细胞透 (CPP) 吸收. 这种方法量化了内部化的完整或降解的,揭示了转位效率.
科学领域:
- 生物化学 生物化学
- 细胞生物学 细胞生物学
- 生物技术是生物技术.
背景情况:
- 细胞透 (CPPs) 通过内细胞分裂和直接转移促进细胞进入.
- 区分这些吸收途径是当前基于细胞的测试具有挑战性的.
- 准确量化CPP内部化路径对于药物输送应用至关重要.
研究的目的:
- 开发一种可靠的方法,以独立区分和量化CPP内细胞和转移.
- 评估结构对转位功效的影响.
- 为了实时分析CPP内部化机制.
主要方法:
- 使用的双标记:DABCYL (暗消火剂) 和碳氧素 (CF,).
- 根据内部化后的质完整性 (完好或退化) 测量光信号的变化.
- 采用了取决于温度的测定方法 (37°C与4°C),以区分转位与内细胞分裂.
主要成果:
- 该试验成功量化了内部化的度和完整性.
- 沸的细胞抑制了酶降解,允许在37°C时评估转位.
- 转位疗效排名:DABCYL-R6-(εCF) K > DABCYL-R4-(εCF) K ≥ CF-R9.9. 转位疗效排名:DABCYL-R6-(εCF) K > DABCYL-R4-(εCF) K ≥ CF-R9. 转位疗效排名: DABCYL-R6-(εCF) K > DABCYL-R4-(εCF) K ≥ CF-R9.
结论:
- 开发的试验可靠地区分和量化CPP吸收途径.
- 这种方法提供了对内部化CPPs的分子状态和位置的洞察.
- 这些发现为优化基于CPP的药物输送系统提供了有价值的工具.
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