Detailed Raman analysis of protein denaturation using vertical flow method and programmable pump

研究成果: Conference contribution同行評審

1 引文 斯高帕斯(Scopus)

摘要

We have developed a vertical flow (VF) method to improve the generation and collection efficiency of Raman scattering signals. The VF method enhances the Raman signal up to 90 times [Li and Hiramatsu, Anal. Chem. 2019, 91, 9806]. The VF method enabled an online Raman measurement of HPLC eluate (LC-Raman) [Lo and Hiramatsu, Anal. Chem. 2020, 92, 14601]. A programmable pump of the HPLC system is available to gradually change the pH value of the solvent by changing the mixing fraction of two solvents. In this study, we used the programmable pump combined with the Raman spectroscopy to probe the denaturation of bovine serum albumin (BSA) change in the range of pH 7-9. The vertical flow method allowed us to record 8300 spectra in 30 min. The obtained 2D Raman data having the spectral and temporal axes was analyzed with singular value decomposition and reconstruction method. We found that four temporal and spectral components predominate the 2D Raman data, namely, (i) the pH-independent Raman signal of BSA, (ii) that due to the denaturation, (iii) the ionization of phosphate in the buffer, and (iv) pH-dependent background. The sufficiently large number of the data points enabled precise determination of the pKa values of the denaturation process of BSA in the range of pH 7-9 and detailed analysis of the structural change in the denaturation.

原文English
主出版物標題Biomedical Vibrational Spectroscopy 2022
主出版物子標題Advances in Research and Industry
編輯Zhiwei Huang
發行者SPIE
ISBN(電子)9781510647855
DOIs
出版狀態Published - 2022
事件Biomedical Vibrational Spectroscopy 2022: Advances in Research and Industry - Virtual, Online
持續時間: 20 2月 202224 2月 2022

出版系列

名字Progress in Biomedical Optics and Imaging - Proceedings of SPIE
11957
ISSN(列印)1605-7422

Conference

ConferenceBiomedical Vibrational Spectroscopy 2022: Advances in Research and Industry
城市Virtual, Online
期間20/02/2224/02/22

指紋

深入研究「Detailed Raman analysis of protein denaturation using vertical flow method and programmable pump」主題。共同形成了獨特的指紋。

引用此