{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T07:55:47Z","timestamp":1791273347807,"version":"4.1.0"},"reference-count":42,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T00:00:00Z","timestamp":1791244800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T00:00:00Z","timestamp":1791244800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Biotech &amp; Bioengineering"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>Vero host cell proteins (HCPs) are critical process\u2010related impurities in Vero cell\u2010derived vaccines and require robust analytical control to support product quality, safety, and manufacturing consistency. During development of the V590 vaccine candidate, discontinuation of the original commercial Vero HCP ELISA prompted evaluation of alternative approaches for HCP monitoring and risk control. We compared first\u2010 and second\u2010generation commercial ELISA kits, assessed the impact of process\u2010representative HCP standards and anti\u2010HCP antibodies on HCP quantitation, and characterized antibody coverage by two\u2010dimensional differential\u2010in\u2010blot electrophoresis. Measured HCP levels in process intermediates and bulk drug substances were strongly influenced by sample composition, HCP standard, antibody reagents, and assay format. ELISA and Simple Western yielded different quantitative results even with the same standard and antibodies, underscoring the assay\u2010dependent nature of HCP measurement. Platform\u2010specific anti\u2010Vero HCP antibodies provided broader coverage of process\u2010representative HCPs than the second\u2010generation commercial ELISA antibodies. Orthogonal characterization by Simple Western and Liquid chromatography\u2010mass spectrometry (LC\u2010MS\/MS) further enabled monitoring of fibronectin, a high\u2010risk Vero HCP that causes filter fouling and compromises downstream process performance. Together, these findings demonstrate that platform\u2010specific immunoassays combined with orthogonal characterization of process\u2010relevant high\u2010risk HCPs provide a robust analytical framework for Vero HCP control and long\u2010term assay lifecycle management in Vero cell\u2010derived vaccine manufacturing.<\/jats:p>","DOI":"10.1002\/bit.70401","type":"journal-article","created":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T07:47:57Z","timestamp":1791272877000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Platform\u2010Specific Immunoassay Development and Orthogonal Characterization for Improved Host Cell Protein Control in Vero Cell\u2010Derived Vaccines"],"prefix":"10.1002","author":[{"given":"Shruti","family":"Patel","sequence":"first","affiliation":[{"name":"Large Molecule Analytical Merck &amp; Co., Inc. Rahway New Jersey USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Young","family":"Zhang","sequence":"additional","affiliation":[{"name":"Large Molecule Analytical Merck &amp; Co., Inc. Rahway New Jersey USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Holly","family":"Guan","sequence":"additional","affiliation":[{"name":"Large Molecule Analytical Merck &amp; Co., Inc. Rahway New Jersey USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emily","family":"Groegler","sequence":"additional","affiliation":[{"name":"Large Molecule Analytical Merck &amp; Co., Inc. Rahway New Jersey USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kuo","family":"Yin","sequence":"additional","affiliation":[{"name":"Large Molecule Analytical Merck &amp; Co., Inc. 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Rahway New Jersey USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2156-9963","authenticated-orcid":false,"given":"Paul F.","family":"Gillespie","sequence":"additional","affiliation":[{"name":"Large Molecule Analytical Merck &amp; Co., Inc. Rahway New Jersey USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yanjie","family":"Wang","sequence":"additional","affiliation":[{"name":"Large Molecule Analytical Merck &amp; Co., Inc. Rahway New Jersey USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dennis","family":"Driscoll","sequence":"additional","affiliation":[{"name":"Large Molecule Analytical Merck &amp; Co., Inc. Rahway New Jersey USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7548-583X","authenticated-orcid":false,"given":"Xuanwen","family":"Li","sequence":"additional","affiliation":[{"name":"Pharmaceutical Analysis &amp; Digital Technologies Merck &amp; Co., Inc. Rahway New Jersey USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8487-5159","authenticated-orcid":false,"given":"Fengqiang","family":"Wang","sequence":"additional","affiliation":[{"name":"Large Molecule Analytical Merck &amp; Co., Inc. Rahway New Jersey USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,10,6]]},"reference":[{"issue":"2","key":"e_1_2_9_2_1","doi-asserted-by":"crossref","first-page":"413","DOI":"10.1002\/bit.26482","article-title":"Comparison of Purification Strategies for Antibodies Used in a Broad Spectrum Host Cell Protein Immunoassay","volume":"115","author":"Baldus P. A.","year":"2018","journal-title":"Biotechnology and Bioengineering"},{"issue":"8","key":"e_1_2_9_3_1","doi-asserted-by":"crossref","first-page":"2082","DOI":"10.1002\/bit.29028","article-title":"CRISPR\u2010Editing of the Vero Cell Line Improves Processability of Live Virus Vaccines by Enabling Targeted Proteolysis of Fibronectin","volume":"122","author":"Cahn J. K. 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