摘要
目的:建立重组人促卵泡激素(rhFSH)N端不均一性的分析方法,并对不同企业样品rh FSH原液的N端不均一性进行比对研究。方法:通过对亚基的还原与烷基化处理方法、糖苷酶切条件及RP-HPLC条件的优化,将完整与N端缺失氨基酸的亚基有效分离,建立了2种rhFSH的N端不均一性分析方法,并利用质谱分别对目的蛋白进行定性鉴别。以国内外企业的rhFSH原液为样品进行N末端不均一性比较。结果:建立的2种RP-HPLC方法均可实现完整与缺失β亚基的有效分离,各样品rhFSH的N端不均一性基本一致。结论:rhFSH国内生物类似药与国外参照药的N端不均一性比较有较高的相似性;本实验建立的2种RP-HPLC方法可评价产品质量,为生物类似药的可比性研究提供技术支持。
Objective： To establish a method for the analysis of N-terminal heterogeneity of recombinant human follicle stimulating hormone （ rhFSH ）, and to compare the N terminal heterogeneity of rhFSH sample solutions from in different enterprises. Methods： Integrated subunit and subunit with N-terminal amino acid residues losing were separated by optimizing the reduction and alkylation process, the glycosidase digestion, and the RP-HPLC conditions. Two methods of analyzing N-terminal heterogeneity of rhFSH were established, and target proteins were further indentified by mass spectrometry. The N-terminal heterogeneity was compared with the domestic and foreign enterprises＇ rhFSH solutions. Results： Integrated 13 subunit and the loss 13 subunit could be separated by two RP-HPLC methods, and the N-terminal heterogeneity of rhFSH from different enterprises were basically consistant. Conclusion： The N-terminal heterogeneity of rhFSH in the domesticbiosimilar druys was of high similarity than that in the international reference drugs ; two established RP-HPLC methods can evaluate the quality of the products and provide technical support for the comparative study on biosimilar.
出处
《药物分析杂志》
CAS
CSCD
北大核心
2017年第5期889-896,共8页
Chinese Journal of Pharmaceutical Analysis
基金
国家科技重大专项--抗体类生物大分子药物质量属性分析标准物质及评价方法研究平台(课题编号:2014ZX09304311)
中检院学科带头人培养基金课题--重组激素类生物类似药质量可比性研究(课题编号:2013X2)
关键词
重组人促卵泡激素
N端不均一性
α、β亚基
可比性研究
： human recombinant follicle stimulating hormone
N-terminal heterogeneity
α, β subunit
comparative research
