Identification of peak 2 protein on right side of main peak of SEC-HPLC in lyophilized human rabies vaccine (Vero cells) stock solution(without human blood albumin)
10.13200/j.cnki.cjb.004260
- VernacularTitle:冻干人用狂犬病疫苗（Vero细胞）原液（无人白）体积分子排阻-高效液相色谱主峰右侧峰2蛋白鉴定
- Author:
ZHANG Qiuju
- Publication Type:Journal Article
- Keywords:
Rabies vaccine(Vero cells)stock solution(without human blood albumin);
Rabies strain 4aGV;
Size exclusion chromatograghy-high performance liquid chromatography(SEC-HPLC);
Protein identification;
nanoLC-MS/MS analysis;
Vaccine virus particle
- From:
Chinese Journal of Biologicals
2024;37(07):801-805
- CountryChina
- Language:Chinese
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Abstract:
Objective To identify the peak 2 protein on the right side of the main peak of size exclusion chromatograghy-high performance liquid chromatography(SEC-HPLC)in freeze-dried human rabies vaccine(Vero cell)stock solution(without human blood albumin),so as to determine the protein composition of the peak and ensure the vaccine quality.Methods The lyophilized human rabies vaccine(Vero cells)stock solution(without human blood albumin)was analyzed by SECHPLC,of which the peak 2 on the right side of the main peak was collected and concentrated by ultrafiltration,then reduced by dithiothreitol(DTT),alkylated by iodoacetamide (IAM) and hydrolyzed by Trypsin. The products were analyzed by nano LC-MS/MS.Results Four proteins were successfully identified in the right peak 2 of SEC-HPLC main peak in lyophilized human rabies vaccine(Vero cell)stock solution(without human blood albumin),and the number of matching peptides for a single protein ranged from 59 to 79. The number of single protein-matching characteristic peptides ranged from 6 to 12. The detection times of single protein-matching characteristic peptide segments ranged from 20 to 36 times. The sequence coverage of the identified proteins ranged from 37. 40% to 64. 31%. A total of 280 peptides participated in the statistics,and the mass spectrum deviation was less than 0. 15 Da.Conclusion The peak 2 on the right side of SEC-HPLC main peak of lyophilized human rabies vaccine(Vero cell)stock solution(without human blood albumin)is derived from rabies strain 4a GV,which is the vaccine particle dissociation.
