使用类似VH的标志性残留物提高单域抗体的溶解度
Yuta Uto1, Makoto Nakakido1,2, Takanori Yokoo1
1Department of Chemistry and Biotechnology, School of Engineering, The University of Tokyo, Tokyo, Japan.
Protein science : a publication of the Protein Society
|June 16, 2025
概括
通过修改标志残留物,通过改造提高溶解度的单域抗体 (sdAbs) 被制造出来. 这些可溶性增强的sdAbs保持了它们的功能,提供了一种改善sdAb生产的方法.
科学领域:
- 生物技术是生物技术.
- 蛋白质工程是指蛋白质工程.
- 免疫学 免疫学 免疫学
背景情况:
- 单域抗体 (sdAbs) 是从重链抗体中衍生出来的,因其小小而受到重视.
- sdAbs 的可溶性受到框架区域 2 (FR2) 中的标志性氨基酸的影响.
- 之前的研究表明,互补性决定的区域也会影响sdAb的溶解性.
研究的目的:
- 设计和提高具有VH类标志残留的低溶解性单域抗体 (sdAbs) 的溶解性.
- 研究特定突变在增强sdAb溶解性和维持功能的作用.
主要方法:
- 在FR2标志残留物中引入了类似VHH的突变,该残留物是两种阿尔帕卡衍生的sdAbs (sdAb A和B).
- 利用空间聚合倾向计算来识别影响溶解性的关键残留物.
- 为sdAb B生成了一个W99A突变,以解决已识别的溶解性问题.
- 使用结构,物理化学和生物物理分析来描述工程 sdAbs.
主要成果:
- 改造后的sdAb A和sdAb B的W99A突变物成功被制备为单体.
- 在FR2中发生的突变改善了sdAb A在没有添加剂的情况下的可溶性.
- 确定W99作为sdAb B低溶性的关键残留物.
- 提高溶解度的sdAbs保留了它们的原始功能.
结论:
- 修改FR2中的标志性残留物可以提高sdAb的溶解度.
- 特定的残留突变,如W99A,可以克服sdAbs.中的可溶性挑战.
- 这种方法可以产生可溶性,功能性的sdabs,即使没有详细的结构信息.
相关概念视频
01:10Antibody Structure
61.4K
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...
61.4K
01:31Hybridoma Technology
15.4K
Hybridoma technology is used for the large-scale production of monoclonal antibodies. Monoclonal antibodies bind to only a single antigenic determinant or epitope. Such antibodies are used in research, diagnostics, and disease therapy. The hybridoma technology established in 1975 by Georges Köhler and Cesar Milstein was awarded the Nobel Prize in Medicine in 1984 for revolutionizing research and therapy.
Hybridoma Selection
Commonly used fusion techniques — electroporation,...
Hybridoma Selection
Commonly used fusion techniques — electroporation,...
15.4K
01:25Antibody Structure and Classes
4.8K
Antibodies, also known as immunoglobulins, are produced by B cells in response to foreign substances, such as bacteria and viruses. These proteins are critical for recognizing and neutralizing these substances, protecting the body from potential harm.
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.
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.
4.8K