Evaluation of removal efficiency of human interferon α2b chromatography process for impurity proteins using liquid chromatography-high resolution mass spectrometry
10.13200/j.cnki.cjb.004420
- VernacularTitle:液相色谱-高分辨质谱法评价人干扰素α2b层析工艺对杂质蛋白的去除效果
- Author:
GUO Ying
- Publication Type:Journal Article
- Keywords:
Human interferon α2b (hIFNα2b);
Related proteins;
Host cell proteins;
Chromatography process;
Liquid chromatography-high resolution mass spectrometry(LC-HRMS)
- From:
Chinese Journal of Biologicals
2025;28(02):179-184
- CountryChina
- Language:Chinese
-
Abstract:
Objective To evaluate the removal efficiency of chromatography process for impurity proteins[IFN-related proteins and host cell proteins (HCPs)]in the purification process of human interferon α2b (h IFNα2b) by liquid chromatogra-phyhigh resolution mass spectrometry (LC-HRMS),in order to provide experimental basis for improving the production process of h IFNα2b.Methods The whole protein levels of IFN-related proteins (including deamidated IFN,oxidized IFN and acetylated IFN) and HCPs during the chromatography process of h IFNα2b were analyzed by LC-HRMS,and the h IFNα2b post-translational modification sites and HCPs were detected on the polypeptide levels.Results HCPs could be removed by hydrophobic interaction chromatography (HIC) M column,HCPs,deamidated IFN and acetylated IFN could be removed by anion exchange chromatography (AEC),and oxidized IFN could be removed by HIC (B column).The major acetylation sites (K34,K121,K131 and K134),major oxidation sites (M16,M21,M59 and M148),as well as Q5 and Q21 of major deamidation sites (N45,N156,Q5,Q21,Q46 and Q124) of h IFNα2b were located on the surface of the protein,which were susceptible to environmental conditions and subjected to acetylation and oxidation.Conclusion In the purification process of h IFNα2b,chromatography purification can effectively remove IFN-related proteins and HCPs,which also indicated that LC-HRMS technology can be used for the research of IFN purification process.
