硫尿素与正常或改性人血清白蛋白结合的表征,使用由捕获制备的亲和力微柱
Saumen Poddar1, Ashley G Woolfork1, Sazia Iftekhar1
1Department of Chemistry, University of Nebraska, Lincoln, NE 68588, USA.
概括
糖尿病诱导的蛋白质修饰,特别是人类血清白蛋白 (HSA) 中的高级糖化最终产品 (AGEs),可以改变药物结合. 这项研究使用微列来显示AGEs增加了对HSA的硫氨酸尿素药物亲和力.
科学领域:
- 生物化学 生物化学
- 药理学 药理学是指药理学的学科.
- 分析化学 分析化学
背景情况:
- 糖尿病可以通过先进的糖化最终产品 (AGEs) 导致蛋白质的修饰.
- 人体血清白蛋白 (HSA) 是一种关键的药物结合蛋白,易受糖 (Go) 和甲基糖 (MGo) 的修饰,形成AGE.
- 由于AGE而改变的HSA结构可能会影响药物相互作用和疗效.
研究的目的:
- 为了研究硫基尿素药物和正常与AGE修饰的HSA.之间的结合相互作用.
- 评估一种新型高性能亲和力微柱技术对评估这些相互作用的有用性.
- 为了量化由AGE修饰引起的药物-HSA结合亲缘关系的变化.
主要方法:
- 使用HSA的非共价捕获来制备高性能亲和力微柱.
- 区域化实验以确定药物保留和结合常数与正常和AGE修饰的HSA (使用Go和MGo).
- 结果与使用不同的HSA固定化技术获得的文献值进行比较.
主要成果:
- 基于捕获的微柱法提供了快速 (3-5分钟) 和精确 (±10-23%) 的全球亲和度常数估计.
- 微柱在长时间使用 (≥60-70注射,1个月) 时表现出极好的稳定性.
- 与正常的HSA相比,AGE修改的HSA对某些硫氨酸urea药物表现出增加的全球亲和度常数 (高达2.1倍).
结论:
- 在微柱中非共价捕获是研究药物-HSA结合的有效和稳定的方法.
- 与糖尿病相关的HSA的AGE修饰显著改变了一些硫基尿素药物的结合亲和力.
- 这种方法为评估临床和研究环境中与改性蛋白质的药物相互作用提供了有价值的工具.
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