Newly developed Laboratory-based Size exclusion chromatography Small-angle x-ray scattering System (La-SSS)

• Published in: Scientific reports 2019-08, Vol.9 (1), p.12610-12, Article 12610
• Main Authors: Inoue, Rintaro, Nakagawa, Tatsuo, Morishima, Ken, Sato, Nobuhiro, Okuda, Aya, Urade, Reiko, Yogo, Rina, Yanaka, Saeko, Yagi-Utsumi, Maho, Kato, Koichi, Omoto, Kazuki, Ito, Kazuki, Sugiyama, Masaaki
• Format: Article
• Language: English
• Subjects: 631/57/2282; 639/925/930/12; 82/16; Biological samples; Cell size; Chromatography; Data acquisition; Flow rates; Humanities and Social Sciences; Ions; Irradiation; Laboratories; multidisciplinary; Science; Science (multidisciplinary); X-ray scattering
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-019-48911-w

To understand a biological system, it is important to observe structures of biomolecules in the solution where the system is functionalized. S mall- A ngle X -ray S cattering coupled with S ize E xclusion C hromatography (SEC-SAXS) is one of techniques to selectively observe the target molecules in the multi-component system. However, this technique is believed to be available only with a synchrotron-based SAXS instrument due to requirement of high beam intensity and, therefore, the limitation of the beam time was obstacle to satisfy demands from many bio-researchers. We newly developed La boratory-based S ize exclusion chromatography S AXS S ystem (La-SSS) by utilizing a latest laboratory-based SAXS instrument and finely optimization of the balance between flow rate, cell volume, irradiation time and so on. La-SSS succeeded not only decoupling of target protein(s) from non-specific aggregates but also measurement of each concerned component in a multi-component system. In addition, an option: “stopping mode”, which is designed for improving statistics of SAXS profile, realized a high S/N data acquisition for the most interesting protein in a multi-component system. Furthermore, by utilizing a column having small bed volume, the small-scale SEC-SAXS study makes available. Through optimization of instrumental parameters and environments, La-SSS is highly applicable for experimental requirements from various biological samples. It is strongly expected that a La-SSS concept must be a normal option for laboratory-based SAXS in the near future.