通过阴离子交换染色学检测到的IgG1中的小形状变化是酸性电荷变体
Masakazu Fukuda1, Melissa A Graewert2, Cy M Jeffries2
1Formulation Development Department, Chugai Pharmaceutical Co., Ltd., 5-5-1 Ukima, Kita-ku, Tokyo, 115-8543, Japan; Laboratory of Functional Molecular Chemistry, Kobe Pharmaceutical University, 4-19-1, Motoyamakita-machi, Higashinada-ku, Kobe, 658-8558, Japan.
治疗性单克隆抗体 (mAb) 酸性变体是构造性的,而不仅仅是翻译后的修饰. 这项研究揭示了芳香区域的局部变性,为mAb的表征提供了新的见解.
科学领域:
- 生物制药特征的表征
- 蛋白质化学 蛋白质化学
- 分析生化学分析生化学
背景情况:
- 在治疗单克隆抗体 (mAb) 充电变体中,特别是酸性变体中,表征翻译后修改是复杂的.
- 现有的方法很难完全阐明这些酸性变体的性质,留下了一个开放的研究领域.
研究的目的:
- 调查治疗mAbs的染色学分离的酸性分数是否存在构造变异.
- 探索使用重新折叠方法生成与形状变异相对应的独特酸性峰值分数的潜力.
主要方法:
- 使用IgG1单克隆抗体 (mAb) 作为案例研究,采用了重新折叠的方法.
- 使用瓜尼丁化变性化mAb,并通过透析与阿尔金因化相对应,逐步重新折叠.
- 使用了分析技术,包括染色学,小角度X射线散射,热分析和光谱学.
主要成果:
- 逐步重新折叠的过程显著增加了IgG1.1.中的酸色谱峰的丰度.
- 这表明,鉴定到的酸性变体主要是形状的,而不是仅仅是翻译后的修饰.
- 微角X射线散射在整个IgG1中没有显著的全球形状变化,这表明局部效应.
结论:
- 治疗mAbs中的酸性变体可以代表构造变体,挑战传统的表征.
- 观察到的形状变化是轻微的和局部的,主要影响芳香氨基酸区域.
- 这项研究为酸性mAb变体的性质提供了关键的见解,改善了它们的表征和理解.
更多相关视频
11:10Antibody Binding Specificity for Kappa (Vκ) Light Chain-containing Human (IgM) Antibodies: Polysialic Acid (PSA) Attached to NCAM as a Case Study
Published on: June 29, 2016
10:50Purification and Analytics of a Monoclonal Antibody from Chinese Hamster Ovary Cells Using an Automated Microbioreactor System
Published on: May 1, 2019
相关概念视频
Antibody Structure
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...
Antibody Structure and Classes
The basic structure of an antibody consists of four protein chains: two identical heavy chains and two identical light chains. These chains are held together by disulfide bonds and other non-covalent interactions, forming a Y-shaped structure.
Two-dimensional Gel Electrophoresis
The first dimension separation uses the isoelectric focusing or IEF technique performed on immobilized pH gradient (IPG) strips that separate proteins according to their isoelectric points.
Biological samples, such...
Ion Exchange
