使用牛津纳米孔技术和双重独特分子标识符的抗体菌体显示选择的深度挖掘
Oscar Mejias-Gomez1, Marta Braghetto1, Morten Kielsgaard Dziegiel Sørensen1
1Department of Biotechnology and Biomedicine, Section for Protein Science and Biotherapeutics, Technical University of Denmark, Kongens Lyngby, Denmark.
New biotechnology
|February 14, 2024
概括
牛津纳米孔技术 (ONT) 使用独特分子标识符 (UMI) 的测序有效监测抗体菌体显示选择. 这种方法量化了多样性,并丰富了罕见的结合物,改善了抗体的发现.
科学领域:
- 生物技术是生物技术.
- 免疫学 免疫学 免疫学
- 分子生物学分子生物学
背景情况:
- 抗体菌体显示对于识别抗体-抗原相互作用至关重要.
- 传统的选方法缺乏对选择丰富性和多样性的洞察力,导致粘合物损失.
研究的目的:
- 引入一种新的下一代测序 (NGS) 方法来监测抗体菌体显示选择.
- 为了使抗体输出选择的深度挖掘,并恢复罕见的结合物.
主要方法:
- 使用牛津纳米孔技术 (ONT) Q20+化学与双重独特分子标识符 (UMI).
- 开发了一个优化的生物信息分析管道,用于监测选拔轮.
- 将ONT-UMI方法与用于抗体查的传统殖民地选择方法进行比较.
主要成果:
- 通过ONT-UMI方法,在整个选择轮中成功监测了多样性和丰富性.
- 能够深度挖掘选择和回收罕见的结合物,即使有>99%丰富的主导结合物.
- 提供了有关亲和力成熟和特异性优化绑定动机的见解.
结论:
- 该ONT-UMI方法提供了传统选的替代方案,量化多样性,并使罕见的粘合剂回收.
- 这个平台支持数据引导的选择策略,以提高抗体的发现.
- 为抗体亲和力成熟和特异性优化提供了洞察力.
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