{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,29]],"date-time":"2026-08-29T07:27:22Z","timestamp":1787988442047,"version":"build-2784847793"},"reference-count":28,"publisher":"MDPI AG","issue":"3","license":[{"start":{"date-parts":[[2025,2,3]],"date-time":"2025-02-03T00:00:00Z","timestamp":1738540800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"State Key Laboratory of Drug Regulatory Science","award":["2023SKLDRS0108"],"award-info":[{"award-number":["2023SKLDRS0108"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Molecules"],"abstract":"<jats:p>PEGylated recombinant human growth hormone (PEG-rhGH) has garnered significant interest in growth hormone research due to its prolonged half-life and improved patient compliance. An accurate evaluation of its biological activity is critical for ensuring the quality of PEG-rhGH-based therapeutics. In this study, we established an in vitro bioactivity assay using a reporter gene method based on the HepG2\/IGF-1 cell line. Key assay parameters, including the initial concentration of PEG-rhGH, serial dilution ratios, cell density, and incubation time, were systematically optimized to generate robust dose\u2013response curves. The assay demonstrated high sensitivity, precision, and reproducibility across multiple batches of PEG-rhGH. The validation results showed an excellent correlation with traditional in vivo animal studies and the Nb2-11 cell proliferation assay, highlighting its suitability for quality control. Furthermore, we developed an ion exchange chromatography (IEC) method to separate five positional isomers of PEG-rhGH, revealing significant differences in bioactivity depending on the PEG modification site. This study demonstrates that the optimized reporter gene assay is not only effective for quality control of PEG-rhGH but also serves as a valuable tool for evaluating and optimizing PEGylated long-acting growth hormone therapeutics.<\/jats:p>","DOI":"10.3390\/molecules30030669","type":"journal-article","created":{"date-parts":[[2025,2,3]],"date-time":"2025-02-03T12:18:56Z","timestamp":1738585136000},"page":"669","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["A Reporter Gene Assay for Measuring the Biological Activity of PEGylated Recombinant Human Growth Hormone"],"prefix":"10.3390","volume":"30","author":[{"ORCID":"https:\/\/orcid.org\/0009-0004-6737-0353","authenticated-orcid":false,"given":"Shaowang","family":"Hu","sequence":"first","affiliation":[{"name":"School of Life Science, Jilin University, Changchun 130012, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiaoming","family":"Zhang","sequence":"additional","affiliation":[{"name":"National Institutes for Food and Drug Control, Beijing 100061, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yi","family":"Li","sequence":"additional","affiliation":[{"name":"National Institutes for Food and Drug Control, Beijing 100061, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jing","family":"Li","sequence":"additional","affiliation":[{"name":"National Institutes for Food and Drug Control, Beijing 100061, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yingwu","family":"Wang","sequence":"additional","affiliation":[{"name":"School of Life Science, Jilin University, Changchun 130012, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8916-6872","authenticated-orcid":false,"given":"Chenggang","family":"Liang","sequence":"additional","affiliation":[{"name":"National Institutes for Food and Drug Control, Beijing 100061, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,2,3]]},"reference":[{"key":"ref_1","first-page":"1141","article-title":"Growth hormone: The expansion of available products and indications","volume":"58","author":"Franklin","year":"2011","journal-title":"Pediatr. 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