一个差异离子移动声学弹射质谱仪系统用于选异构化介导酶药物标
bioRxiv : the preprint server for biology
|January 13, 2025
概括
我们开发了DAEMS,这是一个结合差异离子运动 (DMS) 和声弹射质谱 (AEMS) 的新系统,用于超高吞吐量选. 这种方法可以精确量化代谢异构体,这对于针对病原体的药物发现至关重要.
科学领域:
- 分析化学 分析化学
- 生物化学 生物化学
- 代谢学 代谢学 代谢学
背景情况:
- 代谢学涉及分析众多结构异构体,需要先进的分离技术.
- 目前的方法在同位素分离所需的高分辨率功率上扎.
- 异构酶酶在新陈代谢中至关重要,并代表潜在的药物点.
研究的目的:
- 引入和验证一种新的高通量选系统,DAEMS,将差异离子移动性 (DMS) 与声喷射质谱 (AEMS) 集成在一起.
- 为了证明DAEMS在中间代谢中的结构异构体的同时定量的能力.
- 通过针对病原体特定的异构酶,展示DAEMS作为药物发现的强大工具.
主要方法:
- 开发DAEMS系统,将DMS和AEMS结合起来,用于引入超高吞吐量样本.
- 测量糖酸盐和酸盐/异酸盐同位素对的分辨能力.
- 与致病原体特异性糖酸盐突变酶 (iPGM) 的发光试验对比DAEMS的性能.
- 与其他异构酶 (如 chorismate mutase 和 triosephosphate isomerase) 进行DAEMS的演示.
主要成果:
- 为关键的代谢同位素对实现了1588年和1948年的离子移动性解决功率的记录.
- 在与高通量选相容的速度上成功分离了多个异构酶的基质和产物.
- 在没有内部标准的情况下,证明了结构异构体的同时定量.
- 验证的DAEMS性能与iPGM的发光量测试相比.
结论:
- DAEMS代表了质谱学的重大进步,用于对代谢异构体的高通量选.
- 该系统为复杂的代谢样本提供了前所未有的分辨能力.
- 在各种疾病中,DAEMS为药物发现提供了一个多功能平台,针对异构酶酶.
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