三糖基聚合物用于稳定蛋白质结合物与环境压力因素的稳定
Rock J Mancini1, Juneyoung Lee, Heather D Maynard
1Department of Chemistry and Biochemistry, University of California, Los Angeles, 90095-1569, United States.
Journal of the American Chemical Society
|April 24, 2012
概括
新的三醇侧链聚合物增强了对抗热量和冷化等环境压力因素的蛋白质结合体稳定性. 这些甘油聚合物在保护蛋白质活性方面,与三糖或PEG相比提供了更好的保护.
科学领域:
- 聚合物化学 聚合物化学
- 生物结合的生物结合
- 材料科学 材料科学 材料科学
背景情况:
- 蛋白结合物容易受到环境压力因素的影响,导致活动丧失.
- 稳定策略对于在各种应用中保持蛋白质功能至关重要.
- 醇因其保护性而闻名,但其在聚合物基稳定中的应用需要高效的合成和结合方法.
研究的目的:
- 合成三醇侧链聚合物用于蛋白质稳定.
- 将这些甘油聚合物与蛋白质结合,并评估它们对环境压力因素的稳定性.
- 为了比较这些糖聚合物的有效性与现有的稳定剂,如三和聚乙烯糖醇 (PEG).
主要方法:
- 合成一种新型的甘氨基单体,4,6-O-(4-乙烯基利丁) -α,α-三糖,采用两步过程而无需保护组.
- 通过可逆添加-碎片化链转移 (RAFT) 聚合制备糖聚合物.
- 通过二硫化链接将糖聚合物结合到硫化卵白溶酶.
- 通过冷化和热应力测试评估蛋白质稳定性.
主要成果:
- 精确定义的甘油聚合物,其分子重量在4200至49,500Da之间,已成功合成.
- 与野生类型蛋白质相比,蛋白质-甘油聚合物合物显示出明显增加对冷凍和热的稳定性.
- 在经过十次冷化循环后,Lysozyme保持了高达100%的活性,并在90°C加热1小时后保持了81%的活性.
- 甘油聚合物在稳定蛋白质抵抗冷化和热应激方面表现优于三和PEG.
- 与非共价添加相比,糖聚合物的共价附加提供了优越的冷化稳定性.
结论:
- 三醇侧链聚合物是蛋白质合物的有效稳定剂.
- 开发的甘油聚合物提供了针对冷化和热应激的增强保护,性能优于传统稳定剂.
- 这种方法为改善基于蛋白质的治疗和诊断的保质期和稳定性提供了一个有希望的策略.
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