通过/交换质谱测量探测抗体结构
Zuzana Kalaninová1,2, Lukáš Fojtík1,2, Josef Chmelík3
1BioCeV - Institute of Microbiology of the Czech Academy of Sciences, Vestec, Czech Republic.
Methods in molecular biology (Clifton, N.J.)
|September 4, 2023
概括
/交换 (HDX) 与质谱学 (MS) 结合,为蛋白质结构分析提供了强大的方法. 这种技术对于验证抗体药物等生物制药至关重要,提供了详细的结构洞察力.
科学领域:
- 生物化学 生化学
- 分析化学 分析化学
- 结构生物学 结构生物学
背景情况:
- /交换 (HDX) 与质谱学 (MS) 结合,是蛋白质结构评估的验证技术.
- 监管机构批准HDX-MS用于生物制药验证,特别是针对基于抗体的药物.
- 由于其紧的结构,二硫化键和糖基化,抗体存在分析挑战.
研究的目的:
- 为HDX-MS分析提供一个通用的工作流.
- 强调可靠的蛋白质结构表征的关键步骤.
- 详细优化和空间解析信息的可行性.
主要方法:
- /交换 (HDX) 的实验.
- 质谱仪 (MS) 检测.检测. 质谱仪 (MS) 检测.
- 实现通用工作流程,重点是样本处理和优化.
主要成果:
- 证明一种可靠和详细的蛋白质结构评估方法.
- 对具有挑战性的抗体点的成功应用.
- 确定获得高分辨率结构数据的关键参数.
结论:
- HDX-MS是蛋白质生物制药验证的必不可少的工具.
- 描述的工作流提高了结构信息的可靠性和空间分辨率.
- 优化样品处理和实验参数是成功进行HDX-MS研究的关键.
相关概念视频
Mass Spectrometry: Complex Analysis
820
Mass spectrometry is an important technique for the identification of pure compounds. However, it has some limitations for the analysis of complex mixtures, often due to excessive fragmentation making the spectrum too complicated to decipher. Mass spectrometry can be combined with suitable separation methods in sequence, forming hyphenated methods, which are useful in the analysis of complex mixtures.
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
820
¹H NMR of Labile Protons: Deuterium (²H) Substitution
922
This lesson illustrates the role of deuterium substitution in simplifying the NMR spectrum of compounds comprising labile protons. One method employed is the use of deuterium. Amongst the three isotopes of hydrogen, deuterium (2H) has a nucleus composed of one proton and one neutron. When the D2O solvent is added to a pure dry ethanol solution, its labile proton is substituted with deuterium.
922
High-Resolution Mass Spectrometry (HRMS)
1.4K
The resolution of a mass spectrometer depends on the efficiency of separating ions with different ion masses. The mass of an atom is approximated to the sum of the masses of protons and neutrons inside, considering the masses of protons and neutrons as equal. However, the masses of the proton (1.6726 × 10−24 g) and neutron (1.6749 × 10−24 g) are not truly equal. There is a minor error in the expression of atomic masses relative to the simplest atom of hydrogen. For...
1.4K
Mass Spectrometry: Overview
5.4K
Mass spectrometry is an analytical technique used to determine the molecular mass and molecular formula of a compound. The basic principle of mass spectrometry is to generate ions from the analyte molecule and measure these ion abundances against their molecular mass. One common type of ionization, known as electrospray ionization or EI, bombards the analyte molecules in the gas phase with high-energy electron beams. The electron beams displace an electron from the molecule and leave...
5.4K
Peptide Identification Using Tandem Mass Spectrometry
6.5K
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...
6.5K
Antibody Structure
60.3K
Overview
Antibodies, also known as immunoglobulins (Ig), are essential players of the adaptive immune system. These antigen-binding proteins are produced by B cells and make up 20 percent of the total blood plasma by weight. In mammals, antibodies fall into five different classes, which each elicits a different biological response upon antigen binding.
The Y-Shaped Structure of Antibodies Consists of Four Polypeptide Chains
Antibodies consist of four polypeptide chains: two identical heavy...
Antibodies, also known as immunoglobulins (Ig), are essential players of the adaptive immune system. These antigen-binding proteins are produced by B cells and make up 20 percent of the total blood plasma by weight. In mammals, antibodies fall into five different classes, which each elicits a different biological response upon antigen binding.
The Y-Shaped Structure of Antibodies Consists of Four Polypeptide Chains
Antibodies consist of four polypeptide chains: two identical heavy...
60.3K


