title: Characterization of recombinant human growth hormone variants from sodium hyaluronate-based sustained release formulation of rhGH under heat stress.
abstract: This study provides the findings of investigations of potential product-related variants on recombinant human growth hormone (rhGH) in a once-weekly sustained release formulation (SR-rhGH) of sodium hyaluronate microparticles and on the rhGH bulk solution used as the active ingredient for SR-rhGH under extreme stress conditions of 24 h at 60 °C. The extent of rhGH degradation was much higher in solution (33%) than in SR-rhGH (10%). The degradation products, especiallyMet14 sulfoxide and deamidated rhGH variants, were separated and quantified by a modified reversed-phase high-performance liquid chromatography (RP-HPLC) method at reduced flow rate. The primary degradation product of rhGH was found to be deamidated rhGH, although an unknown peak was also detected. In contrast, the primary degradation product of SR-rhGH was Met14 sulfoxide rhGH, with no unknown peaks. Using a cell proliferation assay, the biological activities of the isolated products of SR-rhGH degradation were found to be equivalent to those of native hGH, as determined by comparison with a National Institute for Biological Standards and Control standard. In conclusion, SR-rhGH is structurally and functionally stable and maintains the intactness of rhGH.
authors.affiliation: Department of Biotechnology, Korea University, Seoul 136-701, South Korea; Biotech Group, LG Life Sciences, Daejeon 305-380, South Korea.
authors.forename: Sun Jin
authors.initials: SJ
authors.lastname: Kim
authors.affiliation: Department of Biotechnology, Korea University, Seoul 136-701, South Korea. Electronic address: cwkim@korea.ac.kr.
authors.forename: Chan Wha
authors.initials: CW
authors.lastname: Kim
mesh_terms: D003692:Delayed-Action Preparations / Q000737:chemistry / Q000493:pharmacokinetics; D006358:Hot Temperature; D019382:Human Growth Hormone / Q000737:chemistry* / Q000493:pharmacokinetics; D006801:Humans; D006820:Hyaluronic Acid / Q000737:chemistry* / Q000493:pharmacokinetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000493:pharmacokinetics
chemical_list: D003692:Delayed-Action Preparations; D011994:Recombinant Proteins; D019382:Human Growth Hormone; D006820:Hyaluronic Acid
keywords: Deamidated form; Human growth hormone; Modified RP–HPLC; Stress test; Sulfoxide form; Sustained release formulation