Resonance energy transfer for assessing the molecular integrity of proteins for local delivery.
Resonance energy transfer for assessing the molecular integrity of proteins for local delivery.
复制标题
共振能量转移用于评估局部递送蛋白质的分子完整性。
DOI：
10.1002/bit.10902
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发表时间：
2004
期刊：
影响因子：
--
通讯作者：
Edelman,ElazerR
中科院分区：
文献类型：
--
作者：
Wu,David;Edelman,ElazerR
目前尚不清楚血管生成生长因子治疗潜力的限制是否源于与递送不足相关的药代动力学问题或靶组织敏感性降低。在这里，我们报告了一种使用共振能量转移来评估局部递送后蛋白质的分子完整性的新方法。例如，我们用荧光供体和非荧光受体对、四甲基罗丹明和 QSY-7 标记成纤维细胞生长因子-2，并在离体牛颈动脉模型中证明该生长因子不受组织施加的蛋白水解限制。我们的数据表明 FGF-2 不太可能在动脉壁内降解，并表明仅药代动力学限制无法完全解释迄今为止在治疗性血管生成中观察到的温和反应。一般来说，共振能量转移可能作为一种新方法来评估局部递送中基于蛋白质的疗法的分子完整性。 © 2004 Wiley 期刊公司版权所有
It remains unclear whether the limitations to the therapeutic potential of angiogenic growth factors stem from pharmacokinetic concerns related to inadequate delivery or from a reduced sensitivity of target tissues. Here, we report a novel method using resonance energy transfer to assess the molecular integrity of proteins after local delivery. As an example, we labeled fibroblast growth factor‐2 with a fluorescent donor and nonfluorescent acceptor pair, tetramethylrhodamine and QSY‐7, and demonstrate in an ex vivo bovine carotid artery model that this growth factor is not limited by proteolytic constraints imposed by the tissue. Our data indicate that FGF‐2 is unlikely to be degraded within the arterial wall and suggest that pharmacokinetic limitations alone cannot fully explain the muted response seen thus far in therapeutic angiogenesis. In general, resonance energy transfer may serve as a novel approach to assess the molecular integrity of protein‐based therapies in local delivery. © 2004 Wiley Periodicals, Inc.