一个轻链错配对双特性的结构研究预测了下游分离的机制
Minjeong Cha1, Ankai Xu2, Ambrose J Williams1
1Department of Purification, Microbiology and Virology, Genentech, Inc, South San Francisco, CA, United States.
Journal of chromatography. A
|July 7, 2024
概括
净化双特异性抗体需要仔细管理配对链. 这项研究表明,错误配对的光链中的电荷和疏水差异如何被利用,以利用色谱进行有效的下游净化.
科学领域:
- 生物技术是生物技术.
- 蛋白质化学 蛋白质化学
- 生物制药制造业 生物制药制造业
背景情况:
- 在单个培养物中组装的双特异性抗体需要精确配对四个蛋白质链 (重型和轻型链).
- 错误的链路配对 (LC不配对) 的可能性使这些复杂的生物制品的下游净化过程复杂化.
- 现有的方法足够解决重链错配,但轻链错配的删除需要更复杂的策略.
研究的目的:
- 调查和报告关于分离轻链错配对的双特异抗体变体的方法.
- 阐明这些错误配对的物种分离的基础结构.
- 为预测净化策略,将蛋白质表面特性与色谱行为相关联.
主要方法:
- 对两种具有轻链不匹配的双特异性抗体变体的案例研究分析.
- 使用阴离子交换和多模态离子交换树脂进行染色分离.
- 分子结构分析和蛋白质表面特性建模.
主要成果:
- 一个带有被破坏的正电荷补丁的轻链错配变体显示,与阴离子交换树脂的结合较弱,使其能够被去除.
- 另一种具有相邻负电荷和疏水性斑块的轻链错配变体对离子交换树脂具有更强的结合,允许选择性保留.
- 蛋白质表面性质的计算建模准确地预测了这两种案例研究中观察到的色谱行为.
结论:
- 双特异抗体的下游净化可以通过利用轻链不匹配引入的电荷和疏水差异来实现.
- 蛋白质表面特性建模是预测和优化复杂抗体格式的净化策略的宝贵工具.
- 这些发现提高了对双特异抗体和类似复杂蛋白疗法的净化方法的理解和应用.
更多相关视频
10:17Creating Highly Specific Chemically Induced Protein Dimerization Systems by Stepwise Phage Selection of a Combinatorial Single-Domain Antibody Library
Published on: January 14, 2020
7.7K
07:22Author Spotlight: Evaluating Biophysical Assays for Characterizing PROTACS Ternary Complexes
Published on: January 12, 2024
3.4K
相关概念视频
00:49Ligand Binding and Linkage
Allosteric proteins have more than one ligand binding site; the binding of a ligand to any of these sites influences the binding of ligands to the other sites. When a protein is allosteric, its binding sites are called coupled or linked. In the case of enzymes, the site that binds to the substrate is known as the active site and the other site is known as the regulatory site. When a ligand binds to the regulatory site, this leads to conformational changes in the protein that can influence the...
02:53Conservative Site-specific Recombination and Phase Variation
Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
The recognition sites for Cre recombinase called LoxP...
The recognition sites for Cre recombinase called LoxP...
00:56Extraction: Advanced Methods
Metal ions can be separated from one another by complexation with organic ligands–the chelating agent– to form uncharged chelates. Here, the chelating agent must contain hydrophobic groups and behave as a weak acid, losing a proton to bind with the metal. Since most organic ligands used in this process are insoluble or undergo oxidation in the aqueous phase, the chelating agent is initially added to the organic phase and extracted into the aqueous phase. The metal-ligand complex is formed in...
01:12Double Resonance Techniques: Overview
Double resonance techniques in Nuclear Magnetic Resonance (NMR) spectroscopy involve the simultaneous application of two different frequencies or radiofrequency pulses to manipulate and observe two distinct nuclear spins. One important application of double resonance is spin decoupling, which selectively suppresses coupling with one type of nucleus while observing the NMR signal from another nucleus, simplifying the spectrum and enhancing resolution.
Spin decoupling is usually achieved by...
Spin decoupling is usually achieved by...