相关实验视频

Updated: Jul 2, 2025

Purification and Analytics of a Monoclonal Antibody from Chinese Hamster Ovary Cells Using an Automated Microbioreactor System

Purification and Analytics of a Monoclonal Antibody from Chinese Hamster Ovary Cells Using an Automated Microbioreactor System

Published on: May 1, 2019

在化条件下使用SEC-MS (dSEC-MS) 进行基于单克隆抗体的重桥形式的深入分析

Rania Benazza¹, Ilias Koutsopetras², Valentine Vaur²

¹Laboratoire de Spectrométrie de Masse BioOrganique, IPHC UMR 7178, Université de Strasbourg, CNRS, 67087 Strasbourg, France; Infrastructure Nationale de Protéomique ProFI-FR2048, 67087 Strasbourg, France.

|February 16, 2024

概括

双硫化物重桥制造出同质的抗体结合体. 与质谱学 (dSEC-MS) 相结合的大小排除染色学精确地描述了这些重桥抗体和Fab片段.

科学领域:

生物结合化学生物结合化学
分析化学分析化学
蛋白质工程是指蛋白质工程.

背景情况:

基于抗体的实体被修改使用二硫化物重新桥接用于新型结合体.
重桥剂的受控还原和共价附着包括每个二硫化物键的有效载荷.
来自不对称的重桥剂的异质物种挑战了传统的SDS-PAGE的特征.

研究的目的:

报告对共价性重桥抗体和Fab片段的深入表征.
为了评估变质化的尺寸排除色谱与质谱学 (dSEC-MS) 结合,用于分析重桥反应.
展示dSEC-MS作为精确质量测量和关键质量属性评估的方法.

主要方法:

抗体和抗原结合碎片 (Fab) 的二硫化物重桥.
降低溶剂可访问的二硫化物键,然后通过重桥剂的共价附着.
使用大小排除染色学与质谱学在变质条件下 (dSEC-MS) 进行表征.

主要成果:

通过dSEC-MS,可以明确识别共价复杂产品和未结合物种.
该方法允许密切监测对结合的trastuzumab和Fab片段的重桥反应.
实现了精确的质量测量和关键质量属性 (CQA) 的评估.

结论:

dSEC-MS是一种强大的工具,用于表征基于抗体的二硫化物重桥式格式.
这种全合一的方法提供了简单的分析和重新弥合效率的确定.
准确的表征对于同质抗体-药物合物 (ADC) 和单克隆抗体 (mAb) 格式的开发至关重要.

关键词:

质谱测量质量谱测量尺寸排除色谱学尺寸排除色谱学

更多相关视频

Simultaneous Affinity Enrichment of Two Post-Translational Modifications for Quantification and Site Localization

Simultaneous Affinity Enrichment of Two Post-Translational Modifications for Quantification and Site Localization

Published on: February 27, 2020

Mass Spectrometric Approaches to Study Protein Structure and Interactions in Lyophilized Powders

Mass Spectrometric Approaches to Study Protein Structure and Interactions in Lyophilized Powders

Published on: April 14, 2015

相关概念视频

Peptide Identification Using Tandem Mass Spectrometry

Peptide Identification Using Tandem Mass Spectrometry

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...