分析化学2024，Vol.52Issue(3)：380-388,中插36-中插40,14.DOI:10.19756/j.issn.0253-3820.231122
基于液相色谱-质谱联用技术的重组人生长激素低丰度产品相关物质结构表征
Structural Characterization of Low Abundance Product-related Substances of Recombinant Human Growth Hormone Using Liquid Chromatography-Mass Spectrometry
刘海龙 1王英武 1冯佳 2张金亮 1王舒越 2刘锋 2张飞飞 2孙丹丹 2焉梦寒 2杨新2
作者信息
- 1. 吉林大学生命科学学院,长春 130023||长春金赛药业有限责任公司,长春 130012
- 2. 长春金赛药业有限责任公司,长春 130012
- 折叠
摘要
Abstract
Recombinant human growth hormone(rhGH)is a protein drug expressed in Escherichia colivia genetic engineering,and plays a vital role in regulating growth,metabolism and energy balance.It is widely used in treatment of various clinical diseases,including growth hormone deficiencies in children and adults.During the development and production process of rhGH,various product-related substances may be formed,potentially impacting the drug's quality.Therefore,detailed structural characterization and rigorous quality control of rhGH and its product-related substances are crucial for ensuring pharmaceutical quality.The European Pharmacopoeia includes methods for quality control of rhGH product-related substances using reversed-phase high-performance liquid chromatography(RP-HPLC)and provides structural identification information for high abundance chromatographic peaks.However,it lacks detailed records for the structural analysis of low abundance product-related substances.In this study,RP-HPLC coupled with liquid chromatography-mass spectrometry(LC-MS)was used to successfully separate and identify three low abundance product-related substances of rhGH.The study revealed that low abundance product-related substances of rhGH generally contained multiple modifications or rare modifications,including double deamidation at N99 and N149,simultaneous loss of two amino acids at the N-terminus and deamidation at N149,and deamidation at Q122,N-terminal formylation,and N-terminal butanoylation.Notably,the discovery of N-terminal butanoylation in rhGH was the first of its kind.This research provided a comprehensive and in-depth characterization of rhGH product-related substances,offering a reliable basis for the development and process improvement of rhGH drugs.关键词
重组人生长激素/结构表征/产品相关物质/翻译后修饰/液相色谱-质谱联用Key words
Recombinant human growth hormone/Structural characterization/Product-related substance/Post-translational modification/Liquid chromatography-mass spectrometry引用本文复制引用
刘海龙,王英武,冯佳,张金亮,王舒越,刘锋,张飞飞,孙丹丹,焉梦寒,杨新..基于液相色谱-质谱联用技术的重组人生长激素低丰度产品相关物质结构表征[J].分析化学,2024,52(3):380-388,中插36-中插40,14.
