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OA北大核心CSTPCD

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.