Identification of different variant strains of inactivated SARS-CoV-2 vaccines(Vero cells) based on UPLC-Q-Orbitrap-MS/MS analysis technology
10.13200/j.cnki.cjb.004549
- VernacularTitle:基于超高效液相色谱-四极杆-静电场轨道阱串联质谱分析技术的不同变异株新型冠状病毒灭活疫苗（Vero细胞）的鉴定
- Author:
YAN Ping
- Publication Type:Journal Article
- Keywords:
Ultra-performance liquid chromatography-quadrupole-electrostatic field orbitrap tandem mass spectrometry(UPLC-Q-Orbitrap-MS/MS);
SARS-CoV-2 vaccine;
Peptide sequences;
Identification
- From:
Chinese Journal of Biologicals
2025;38(08):944-950
- CountryChina
- Language:Chinese
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Abstract:
Objective Toestablishanultra-performanceliquidchromatography-quadrupole-electrostaticfieldorbitraptandem mass spectrometry(UPLC-Q-Orbitrap-MS/MS) analysis technology for the identification of different variants of inactivated SARS-CoV-2 vaccines(Vero cells), so as to provide a new evaluation direction for vaccine supervision.Methods Peptide samples were obtained from inactivated SARS-CoV-2 vaccines after reductive alkylation, tryptic digestion, and desalting of SDB-RPS desalting column, and then were eluted by CDS C18 column(0. 075 mm × 250 mm × 1. 9 μm) in gradient program, with 0. 1% formic acid and 3% dimethyl sulfoxide as mobile phase A, 0. 1% formic acid, 3% dimethyl sulfoxide and 80% acetonitrile as mobile phase B. The column temperature was 50 ℃, the flow rate was 300 nL/min, and the injection volume was 5 μL. Then UPLC-Q-Orbitrap-MS/MS analysis technology was established, the mass spectrometry detection was scanned by data-dependent acquisition(DDA) mode, and the sample peptide information was retrieved and analyzed by MaxQuant(V 2.4.2.0) software.Results The established method was used to analyze and determine the inactivated SARSCoV-2 vaccines(Vero cells) of different variants, and the conserved peptides and characteristic peptide sequences of S protein and N protein of different variants of SARS-CoV-2 were screened and extracted, and the peptide mass-to-charge ratio(m/z) was combined to obtain the specific protein database of different variants. By comparing the peptide information detected by the mass spectrometry of the sample, it is possible to identify different variants of inactivated SARS-CoV-2vaccines(Vero cells).Conclusion The UPLC-Q-Orbitrap-MS/MS analysis technology established in this study has the advantages of high sensitivity, rapidity and strong specificity, which can be effectively used for the identification and screening of strains of inactivated SARS-CoV-2 vaccines(Vero cells), can provide a technical reference for the identification of SARS-CoV-2 species in other viral vaccines or biological samples, and provide a new perspective for the quality and safety supervision of vaccines.
