清洁和完整的蛋白质消化与自解耐药的素为片映射
Beatrice Muriithi1, Samantha Ippoliti1, Abraham Finny1
1Waters Corporation, 34 Maple Street, Milford, Massachusetts 01757, United States.
Journal of proteome research
|October 11, 2024
概括
化学修饰的素抵抗自分解,改善了单克隆抗体的类映射. 这种增强的酶提供了更大的稳定性和消化效率,减少了错过的裂,以获得可靠的生物治疗特征.
科学领域:
- 生物化学 生化学
- 分析化学 分析化学
- 生物技术是生物技术.
背景情况:
- 片映射对于识别和量化蛋白质特异性片至关重要.
- 素自解降低了切割的特异性和有效性在基映射.
- 生物治疗单克隆抗体的表征需要可靠的类映射方法.
研究的目的:
- 报告生物治疗单克隆抗体的高效和可重现的映射.
- 为了突出同质化学修饰的素的特性.
- 在抗体表征中证明耐自解素的实用性.
主要方法:
- 使用融温度和展开能量,描述化学修饰的素的热稳定性.
- 通过在消化后测量完整质量峰值面积来评估自身溶解阻力.
- 使用LC-MS进行酶强度和分子量分布的比较.
- 在单克隆抗体基映射中错过的切割减少的评估.
主要成果:
- 化学修饰的素显示化温度增加了54%,展开能量增加了84%.
- 保持了90%以上的完整质量峰值面积,证实了自身溶解耐药性.
- 与其他试类型相比,改性试呈现了10倍的强度和更窄的分子量分布.
- 在30分钟的消化过程中,经过修改的素观察到>30%的错过裂减少.
结论:
- 均的化学修饰素提供了卓越的热稳定性和自身溶解耐受性.
- 这种修改后的素可以使单克隆抗体的一致且可靠的映射成为可能.
- 增强的酶提高了消化效率,减少了生物治疗特征的错过裂.
相关概念视频
01:02Protein Digestion
102.9K
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.
102.9K
01:33Peptide Identification Using Tandem Mass Spectrometry
6.4K
Tandem mass spectrometry, also known as MS/MS or MS2, is an analytical technique that employs two mass analyzers. Essentially it is a series of mass spectrometers that helps isolate a particular biomolecule and then helps study its chemical properties.
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
This technique helps gather information regarding the protein from which the peptide was obtained and to study the peptides’ amino acid sequence. Identifying peptides from a complex mixture is an important component of the growing field of...
6.4K