NISTの新手法、ウイルスのトゲトゲ兵器に付着した糖の同定が可能に(New NIST Method Can Better Identify Sneaky Sugars on Viruses’ Spiky Weapons)


2023-09-28 米国国立標準技術研究所(NIST)



SARS-CoV-2スパイクタンパク質の部位特異的糖鎖プロファイルを複数のソースから決定する。 Determining Site-Specific Glycan Profiles of Recombinant SARS-CoV-2 Spike Proteins from Multiple Sources

Meghan C. Burke,Yi Liu, Concepcion Remoroza, Yuri A. Mirokhin, Sergey L. Sheetlin, Dmitrii V. Tchekhovskoi, Guanghui Wang, Xiaoyu Yang, and Stephen E. Stein
Journal of Proteome Research  Published:August 30, 2023


Abstract Image

Glycopeptide Abundance Distribution Spectra (GADS) were recently introduced as a means of representing, storing, and comparing glycan profiles of intact glycopeptides. Here, using that representation, an extensive analysis is made of multiple commercial sources of the recombinant SARS-CoV-2 spike protein, each containing 22 N-linked glycan sites (sequons). Multiple proteases are used along with variable energy fragmentation followed by ion trap confirmation. This enables a detailed examination of the reproducibility of the method across multiple types of variability. These results show that GADS are consistent between replicates and laboratories for sufficiently abundant glycopeptides. Derived GADS enable the examination and comparison of the glycan profiles between commercial sources of the spike protein. Multiple distinct glycopeptide distributions, generated by multiple proteases, confirm these profiles. Comparisons of GADS derived from 11 sources of recombinant spike protein reveal that sources for which protein expression methods were the same produced near-identical glycan profiles, thereby demonstrating the ability of this method to measure GADS of sufficient reliability to distinguish different glycoform distributions between commercial vendors and potentially to reliably determine and compare differences in glycosylation for any glycoprotein under different conditions of production. All mass spectrometry data files have been deposited in the MassIVE repository under the identifier MSV000091776.