Department of Oncology, Georgetown University, Washington, DC, 20057, USA.
Innovation Center for Biomedical Informatics (ICBI), Georgetown University, Washington, DC, 20057, USA.
BMC Mol Cell Biol. 2021 Mar 6;22(1):17. doi: 10.1186/s12860-021-00354-w.
Surface plasmon resonance is a label-free biophysical technique that is widely used in investigating biomolecular interactions, including protein-protein, protein-DNA, and protein-small molecule binding. Surface plasmon resonance is a very powerful tool in different stages of small molecule drug development and antibody characterization. Both academic institutions and pharmaceutical industry extensively utilize this method for screening and validation studies involving direct molecular interactions. In most applications of the surface plasmon resonance technology, one of the studied molecules is immobilized on a microchip, while the second molecule is delivered through a microfluidic system over the immobilized molecules. Changes in total mass on the chip surface is recorded in real time as an indicator of the molecular interactions.
Quality and accuracy of the surface plasmon resonance data depend on experimental variables, including buffer composition, type of sensor chip, coupling chemistry of molecules on the sensor surface, and surface regeneration conditions. These technical details are generally included in materials and methods sections of published manuscripts and are not easily accessible using the common internet browser search engines or PubMed. Herein, we introduce a surface plasmon resonance database, www.sprdatabase.info that contains technical details extracted from 5140 publications with surface plasmon resonance data. We also provide an analysis of experimental conditions preferred by different laboratories. These experimental variables can be searched within the database and help future users of this technology to design better experiments.
Amine coupling and CM5 chips were the most common methods used for immobilizing proteins in surface plasmon resonance experiments. However, number of different chips, capture methods and buffer conditions were used by multiple investigators. We predict that the database will significantly help the scientific community using this technology and hope that users will provide feedback to improve and expand the database indefinitely. Publicly available information in the database can save a great amount of time and resources by assisting initial optimization and troubleshooting of surface plasmon resonance experiments.
表面等离子体共振是一种无标记的生物物理技术,广泛用于研究生物分子相互作用,包括蛋白质-蛋白质、蛋白质-DNA 和蛋白质-小分子结合。表面等离子体共振是小分子药物开发和抗体表征的各个阶段的非常强大的工具。学术机构和制药行业都广泛使用这种方法进行涉及直接分子相互作用的筛选和验证研究。在表面等离子体共振技术的大多数应用中,研究的分子之一被固定在微芯片上,而第二个分子通过微流控系统输送到固定分子上。芯片表面上总质量的变化被实时记录,作为分子相互作用的指标。
表面等离子体共振数据的质量和准确性取决于实验变量,包括缓冲液组成、传感器芯片类型、传感器表面分子的偶联化学和表面再生条件。这些技术细节通常包含在已发表的手稿的材料和方法部分中,并且使用常见的互联网浏览器搜索引擎或 PubMed 不易访问。在此,我们介绍了一个表面等离子体共振数据库,www.sprdatabase.info,其中包含从 5140 篇具有表面等离子体共振数据的出版物中提取的技术细节。我们还提供了不同实验室偏好的实验条件分析。这些实验变量可以在数据库中搜索,并帮助该技术的未来用户设计更好的实验。
胺偶联和 CM5 芯片是在表面等离子体共振实验中固定蛋白质最常用的方法。然而,许多不同的芯片、捕获方法和缓冲条件被多个研究人员使用。我们预测该数据库将极大地帮助使用该技术的科学界,希望用户提供反馈信息以不断改进和扩展数据库。数据库中公开的信息可以通过协助表面等离子体共振实验的初始优化和故障排除,节省大量的时间和资源。