Establishment and verification of ultra high performance liquid chromatography and tandem mass spectrometry for determination of receptor binding domain protein content of virus-like particle SARS-CoV-2 vaccine
10.13200/j.cnki.cjb.004583
- VernacularTitle:病毒样颗粒新冠疫苗受体结合结构域蛋白含量超高效液相色谱-串联质谱检测方法的建立及验证
- Author:
GONG Liping
- Publication Type:Journal Article
- Keywords:
Ultra high performance liquid chromatography and tandem mass spectrometry(UHPLC-MS/MS);
SARS-CoV-2 vaccine;
Receptor binding domain(RBD);
Characteristic peptide
- From:
Chinese Journal of Biologicals
2025;38(10):1211-1218+1225
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish an ultra high performance liquid chromatography and tandem mass spectrometry(UHPLCMS/MS) method for determination of the protein content of the receptor binding domain(RBD) of virus-like particle(VLP)SARS-CoV-2 vaccine, and to validate and preliminarily apply it, so as to provide technical support for the quality control.Methods The characteristic peptide of RBD protein of SARS-CoV-2 vaccine was screened by high resolution mass spectrometer, and the content of RBD protein was determined by UHPLC-MS/MS. The separation was performed on a Thermo Fisher Hypersil GOLD C18(100 mm × 2. 1 mm, 3 μm) using the mobile phase A consisting of 0. 1% formic acid aqueous solution and mobile phase B consisting of 0. 1% acetonitrile formate solution at a flow rate of 0. 3 mL/min, via the gradient elution,0-3 min,2% B;3-10 min,2%→50% B;10-10.1 min,50%→90% B;10.1-12 min,90% B;12-12.1 min,90%→2% B; 12. 1-15 min, 2% B. The column temperature was 40 ℃ and the sample size was 1 μL. The specificity, stability,precision, linearity, limit of quantitation(LOQ), recovery and repeatability of the method were verified. Six batches of VLP SARS-CoV-2 vaccine bulk solution and six batches of finished products were detected by the method, and the correlation between the method and the method by amino acid analyzer was analyzed.Results The blank solution had no interference with the determination of characteristic peptide content. The test product solution showed good stability within 24 h at room temperature. The relative standard deviation(RSD) of the peak area of the characteristic peptide was 1. 0% after six consecutive injections of the characteristic peptide reference solution. The standard curve had a good linear relationship at the characteristic peptide concentration ranged from 0. 008 954 to 0. 358 200 μg/mL, and the LOQ was 0. 008 954 μg/mL. The RSDs of recovery rates of characteristic peptide in bulk solution and vaccine samples were all less than 10%. The contents of six batches of vaccine bulk solution and six batches of vaccine samples were 0. 169 40-0. 196 60 mg/mL and 0. 014 35-0. 020 50 mg/mL, respectively. The deviation of RBD protein content in VLP SARS-CoV-2 vaccine bulk solution determined by amino acid analyzer and characteristic peptide method was 5. 3%, and the determination results were basically the same.Conclusion The established UHPLC-MS/MS method is simple to operate with strong specificity and accurate quantitation,and can be used for the determination of RBD protein content in VLP SARS-CoV-2 vaccine.
