用LC-MS/MS对蛋白质消化和固体相提取策略进行比较评估,用于使用LC-MS/MS在血中的寡核酸量化
Vidya Annavarapu1, Michael G Bartlett1
1Department of Pharmaceutical and Biomedical Sciences, College of Pharmacy, University of Georgia, Athens, GA 30602-2352, USA.
Journal of chromatography. A
|January 20, 2026
概括
本研究探讨了使用固相提取 (SPE) 来提取寡核酸的新型非表面活性缓冲剂. 新方法提供了更容易的样本制备和良好的回收,用于核酸治疗药物开发.
科学领域:
- 生物化学 生物化学
- 分析化学 分析化学
- 制药科学 制药科学
背景情况:
- 氧核酸治疗药物由于极性和核酶易感性而存在提取和量化挑战.
- 固相提取 (SPE) 是敏感和选择性寡核酸分析的关键技术.
- 针对SPE的传统表面活性剂基缓冲剂正在重新评估,以改进方法.
研究的目的:
- 为了研究一个非表面活性剂缓冲器预处理模块的寡核酸SPE.
- 为了评估这个模块的性能,使用基于WAX和混合模式的SPE微板.
- 验证SPE方法并将其应用于体外稳定性研究.
主要方法:
- 使用SPE的非表面活性剂缓冲器预处理模块.
- 采用基于WAX的和混合模式的SPE微板.
- 验证了SPE方法并评估了寡核酸回收和样本准备的方便性.
主要成果:
- 非表面活性剂缓冲剂在基于WAX的和混合模式的SPE微板上表现出良好的回收和简化样品制备.
- 传统的以蛋白酶K为基础的表面活性剂缓冲剂在基于WAX的SPE微板上显示出改善的恢复.
- 开发的方法已经成功验证并应用于体外稳定性研究.
结论:
- 非表面活性剂缓冲器预处理模块是一种可行的,高效的替代品,可以替代寡核酸SPE.
- 基于WAX的和混合模式的SPE微板都适合使用这种新方法提取寡核酸.
- 进一步优化可能涉及将表面活性剂基缓冲器与特定的SPE格式相结合,以提高回收率.
相关概念视频
Protein Digestion
110.6K
Protein digestion begins in the stomach, where the highly acidic environment can easily disrupt protein structure by exposing the peptide bonds of polypeptide chains. After polypeptide chains are broken into individual amino acids by a series of digestive enzymes, the amino acids are transported to the liver via the bloodstream to produce energy.
110.6K
Cephalic Phase of Digestion
3.6K
The process of digestion is composed of three stages – cephalic, gastric, and intestinal – each with a distinct control center. The cephalic phase is the first stage, and it starts even before the food enters the stomach. It is controlled by the central nervous system and is initiated by any food-related sensory stimuli, such as the sight and smell of food, which send signals to the brain. While eating, the taste receptors intensify these signals, which travel to the cerebral cortex...
3.6K
Intestinal Phase of Digestion
7.1K
The intestinal phase of digestion is the third and final stage of the digestive process, occurring after the cephalic and gastric phases. It begins when chyme, a partially digested mixture of food and digestive enzymes, enters the small intestine from the stomach. This phase is crucial for nutrient absorption and involves complex hormonal and enzymatic interactions.
The arrival of the chyme in the small intestine distends the duodenum, which triggers the enterogastric reflex. This distension...
The arrival of the chyme in the small intestine distends the duodenum, which triggers the enterogastric reflex. This distension...
7.1K
Gastric Phase of Digestion
3.1K
The gastric phase of digestion begins as soon as food enters the stomach. The incoming food bolus triggers neural and hormonal mechanisms, which last approximately 3 to 4 hours. During this phase, the stomach undergoes significant changes to prepare the food for further digestion and absorption.
When food enters the stomach, it stretches the stomach walls and activates stretch receptors. This triggers local reflexes of the enteric nervous system, mediated through the myenteric plexus. These...
When food enters the stomach, it stretches the stomach walls and activates stretch receptors. This triggers local reflexes of the enteric nervous system, mediated through the myenteric plexus. These...
3.1K
What is Monogastric Digestion?
75.1K
The human body contains a monogastric digestive system. In a monogastric digestive system, the stomach only contains one chamber in which it digests food. Several other animal species also have monogastric digestive systems, including pigs, horses, dogs, and birds. This chapter, however, focuses on the human digestive system.
75.1K
Phase Diagrams
49.0K
A phase diagram combines plots of pressure versus temperature for the liquid-gas, solid-liquid, and solid-gas phase-transition equilibria of a substance. These diagrams indicate the physical states that exist under specific conditions of pressure and temperature and also provide the pressure dependence of the phase-transition temperatures (melting points, sublimation points, boiling points). Regions or areas labeled solid, liquid, and gas represent single phases, while lines or curves represent...
49.0K


