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相关概念视频

Electrospray Ionization (ESI) Mass Spectrometry01:12

Electrospray Ionization (ESI) Mass Spectrometry

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Higher molecular weight biomolecules are nonvolatile compounds that may decompose before ionizing or vaporizing during mass analysis with conventional electron impact ionization methods. Accordingly, electrospray ionization (ESI) is the favored method for vaporizing and ionizing biomolecules as it circumvents rapid fragmentation and enables the recording of mass signals for the entire biomolecule.
ESI utilizes electrical energy to transfer ions from the liquid phase of the sample into the...
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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...
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Selected Reaction Monitoring Mass Spectrometry for Absolute Protein Quantification
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基于校准粒子计数的绝对蛋白质量定量,使用电子喷雾差分移动性分析.

Wei Mi1, Xinyi Zhang1, Bin Wang1

  • 1National Institute of Metrology, No.18 Beisanhuan Donglu, Beijing, 100029, China.

Analytica chimica acta
|April 18, 2024
PubMed
概括

精确的蛋白质定量对于诊断和药物至关重要. 本研究引入了一种使用电喷射差移动性分析 (ES-DMA) 的校准颗粒计数方法,以及用于精确测量蛋白质的凝结颗粒计数器 (CPC).

关键词:
检测效率的检测效率是什么电子喷雾-差分移动性分析-凝结粒子计数器粒子计数 粒子计数 粒子计数蛋白质量化量化的蛋白质.

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科学领域:

  • 生物技术是生物技术.
  • 分析化学 分析化学
  • 生物物理学的生物物理.

背景情况:

  • 精确的蛋白质量化对于体外诊断和药物产品的可追溯性至关重要.
  • 现有的方法可能会面临各种蛋白质大小和缺乏参考标准的挑战.

研究的目的:

  • 开发一种使用校准粒子计数的蛋白质绝对量化方法.
  • 用经过认证的参考材料和各种蛋白质类型验证方法的性能.

主要方法:

  • 使用电喷差移动性分析 (ES-DMA) 与凝结颗粒计数器 (CPC) 结合用于校准颗粒计数.
  • 使用校准仪确定检测效率,使用牛血清白蛋白 (BSA) 和NIMCmAb参考材料验证性能.
  • 研究了不同蛋白质的检测效率和粒子大小分布之间的线性关系.

主要成果:

  • 通过ES-DMA-CPC方法,可以根据测量颗粒数量和确定检测效率来绝对量化蛋白质.
  • 使用认证的参考材料 (BSA,NIMCmAb) 验证了测量性能.
  • 在蛋白质检测效率和颗粒大小之间观察到一个线性关系,适用于各种蛋白质,包括肌球蛋白,纤维素原,NIST参考材料8671,D-二次体和谷氨S转移酶二次体.

结论:

  • 校准粒子计数方法为蛋白质定量提供了一个快速,准确和非破坏性的方法.
  • 这种方法对于测量不同大小的蛋白质颗粒特别有效,即使没有特定的参考标准.