{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,13]],"date-time":"2026-08-13T00:11:45Z","timestamp":1786579905563,"version":"build-2736575974"},"reference-count":38,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2024,8,29]],"date-time":"2024-08-29T00:00:00Z","timestamp":1724889600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Merck KGaA (Darmstadt, Germany)"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Highly purified human menopausal gonadotropin (HP-hMG [Menopur\u00ae, Ferring Pharmaceuticals, Saint-Prex, Switzerland]) contains a 1:1 ratio of follicle-stimulating hormone (FSH) and luteinizing hormone (LH). This analysis aimed to assess gonadotropin (FSH, LH and hCG) abundance in HP-hMG and clarify the source of hCG by assessing the presence of sulfated glycans, which are diagnostic for pituitary hCG forms due to their distinct glycosylation patterns. Additionally, the purity of each sample, their specific components, and their oxidation levels were assessed. HP-hMG samples (three of Menopur\u00ae and two of Menogon\u00ae Ferring Pharmaceuticals, Saint-Prex, Switzerland) were included in the current analyses. Brevactid\u00ae (urinary hCG; Ferring Pharmaceuticals, Saint-Prex, Switzerland) and Ovidrel\u00ae (recombinant hCG; Merck KGaA, Darmstadt, Germany) were used as control samples. Glycopeptide mapping and analysis of impurities were carried out by liquid chromatography\u2013tandem mass spectrometry (LC-MS\/MS). Oxidation was assessed through reducing peptide mapping using LC-MS\/MS. The FSH and LH in the HP-hMG samples showed sulfated glycans, while no signals of sulfated glycopeptides were detected on any site of the beta subunit of hCG. HP-hMG test samples presented the same hCG glycan distribution as the control sample (placental hCG, Brevactid\u00ae) extracted from the urine of pregnant women, suggesting a non-pituitary source of hCG. Protein impurities were estimated to constitute approximately 20\u201330% of the entire HP-hMG protein content in the test samples. More than 200 non-gonadotropin proteins were identified in the HP-hMG test samples, of which several were involved in embryonic development or pregnancy. The alpha subunit of the tested samples was strongly oxidized, with a relative abundance of 20% of the total gonadotropin content. Without taking into account all the protein impurities, the beta subunit of LH was detected only in traces (0.9\u20131.2%) in all tested HP-HMG samples, confirming the data obtained by intact molecule analysis, while high levels of beta hCG (18\u201347%) were observed. Advanced molecular analysis of HP-hMG indicates a primarily placental origin of hCG, as evidenced by the absence of hCG sulfated glycans and the predominance of placental non-sulfated hCG in LH activity. The analysis revealed 20\u201330% of protein impurities and a significant presence of oxidized forms in the HP-hMG samples. These findings are critical for understanding the quality, safety, and clinical profile of HP-hMG.<\/jats:p>","DOI":"10.3390\/ijms25179405","type":"journal-article","created":{"date-parts":[[2024,8,29]],"date-time":"2024-08-29T11:33:37Z","timestamp":1724931217000},"page":"9405","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Analytical Investigation of the Profile of Human Chorionic Gonadotropin in Highly Purified Human Menopausal Gonadotrophin Preparations"],"prefix":"10.3390","volume":"25","author":[{"given":"Angela","family":"Capolupo","sequence":"first","affiliation":[{"name":"Characterization & Innovative Analytics Unit\u2014Analytical Development Biotech\u2014Global Analytical Development\u2014Global Development & Launch\u2014Global Healthcare Operation, Merck Serono S.p.A., 00012 Guidonia, Italy, an affiliate of Merck KGaA, Darmstadt, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sofia","family":"Petrocchi","sequence":"additional","affiliation":[{"name":"Characterization & Innovative Analytics Unit\u2014Analytical Development Biotech\u2014Global Analytical Development\u2014Global Development & Launch\u2014Global Healthcare Operation, Merck Serono S.p.A., 00012 Guidonia, Italy, an affiliate of Merck KGaA, Darmstadt, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maura","family":"Melchiorre","sequence":"additional","affiliation":[{"name":"Characterization & Innovative Analytics Unit\u2014Analytical Development Biotech\u2014Global Analytical Development\u2014Global Development & Launch\u2014Global Healthcare Operation, Merck Serono S.p.A., 00012 Guidonia, Italy, an affiliate of Merck KGaA, Darmstadt, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kim","family":"Jonas","sequence":"additional","affiliation":[{"name":"Department of Women and Children\u2019s Health, School of Life Course and Population Health Sciences, King\u2019s College London, London WC2R 2LS, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"D\u2019Hooghe","sequence":"additional","affiliation":[{"name":"Global Medical Affairs\u2014Fertility, Merck KGaA, 64293 Darmstadt, Germany"},{"name":"Department of Development and Regeneration, Laboratory of Endometrium, Endometriosis & Reproductive Medicine, KU Leuven, 3000 Leuven, Belgium"},{"name":"Department of Obstetrics, Gynecology, and Reproductive Sciences, Yale University Medical School, New Haven, CT 06520-8065, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4206-737X","authenticated-orcid":false,"given":"Aylin","family":"Hanyaloglu","sequence":"additional","affiliation":[{"name":"Department of Metabolism, Digestion and Reproduction, Imperial College, London SW7 2AZ, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sesh","family":"Sunkara","sequence":"additional","affiliation":[{"name":"Department of Women and Children\u2019s Health, School of Life Course and Population Health Sciences, King\u2019s College London, London WC2R 2LS, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Angelo","family":"Palmese","sequence":"additional","affiliation":[{"name":"Characterization & Innovative Analytics Unit\u2014Analytical Development Biotech\u2014Global Analytical Development\u2014Global Development & Launch\u2014Global Healthcare Operation, Merck Serono S.p.A., 00012 Guidonia, Italy, an affiliate of Merck KGaA, Darmstadt, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-0899-7826","authenticated-orcid":false,"given":"Beste","family":"Ozgumus","sequence":"additional","affiliation":[{"name":"Department of Chemical Sciences, University of Naples Federico II, Via Cintia, 80126 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Angela","family":"Amoresano","sequence":"additional","affiliation":[{"name":"Department of Chemical Sciences, University of Naples Federico II, Via Cintia, 80126 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Gabriella","family":"Angiuoni","sequence":"additional","affiliation":[{"name":"Characterization & Innovative Analytics Unit\u2014Analytical Development Biotech\u2014Global Analytical Development\u2014Global Development & Launch\u2014Global Healthcare Operation, Merck Serono S.p.A., 00012 Guidonia, Italy, an affiliate of Merck KGaA, Darmstadt, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1220-410X","authenticated-orcid":false,"given":"Susana","family":"Montenegro","sequence":"additional","affiliation":[{"name":"Global Medical Affairs\u2014Fertility, Merck KGaA, 64293 Darmstadt, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrizia","family":"Simone","sequence":"additional","affiliation":[{"name":"Characterization & Innovative Analytics Unit\u2014Analytical Development Biotech\u2014Global Analytical Development\u2014Global Development & Launch\u2014Global Healthcare Operation, Merck Serono S.p.A., 00012 Guidonia, Italy, an affiliate of Merck KGaA, Darmstadt, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9920-3208","authenticated-orcid":false,"given":"Monica","family":"Lispi","sequence":"additional","affiliation":[{"name":"Global Medical Affairs\u2014Fertility, Merck KGaA, 64293 Darmstadt, Germany"},{"name":"Unit of Endocrinology, International PhD School in Clinical and Experimental Medicine, University of Modena and Reggio Emilia, 41121 Modena, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,8,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","unstructured":"Lunenfeld, B., Bilger, W., Longobardi, S., Alam, V., D\u2019Hooghe, T., and Sunkara, S.K. 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