Chromatographic purification of recombinant group B meningococcal fHbp protein and optimization of endotoxin removal process
10.13200/j.cnki.cjb.004787
- VernacularTitle:重组B群脑膜炎球菌H因子结合蛋白的层析纯化及内毒素去除工艺优化
- Author:
Yuxuan ZHAO
- Publication Type:Journal Article
- Keywords:
Group B Neisseria meningitidis;
Factor H-binding protein(fHbp);
Endotoxin;
Chromatography
- From:
Chinese Journal of Biologicals
2026;39(10):1198-1204+1212
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish an efficient purification and endotoxin [lipopolysaccharide(LPS)] removal process for group B Neisseria meningitidis V2.16 type factor H-binding protein(fHbp), obtain recombinant fHbp protein with high purity and low endotoxin, and provide technical support for the development of related vaccines.Methods The fHbp protein was expressed in E. coli BL21(DE3) mediated by major outer membrane lipoprotein(MLP) signal peptide. After bacterial cell disruption and inclusion body denaturation-renaturation, the LPS removal effects of different types of ceramic hydroxyapatite(CHT)(CHT TypeⅠ/Ⅱ/XT), cation exchange resin(Mab 60 SP) and anion exchange resin(Capto Q ImpRes) were compared under different chromatography modes(flow-through, bind-elution) and process parameters. Three batches of parallel verification experiments were carried out by combining the optimal chromatography methods to optimize the process conditions.Results The optimal chromatography combination process was determined as follows: CHT Type XT resin in flow-through mode, Capto Q ImpRes resin in bind-elution mode, Mab 60 SP resin in bind-elution mode. In three batches of parallel experiments, the average total LPS removal rate was 94. 45% with the highest of 95. 21%, and the protein purity reached over95%.Conclusion The three-step chromatography combination process can efficiently remove endotoxin from recombinant fHbp protein with good repeatability, providing a reliable technical basis for the industrial production of group B Neisseria meningitidis recombinant protein vaccines.
