{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,25]],"date-time":"2026-04-25T07:33:21Z","timestamp":1777102401583,"version":"3.51.4"},"reference-count":32,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2025,10,26]],"date-time":"2025-10-26T00:00:00Z","timestamp":1761436800000},"content-version":"vor","delay-in-days":25,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biotechnology Journal"],"published-print":{"date-parts":[[2025,10]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>Proteolytic degradation of recombinant proteins in Chinese hamster ovary (CHO) cells remains a major challenge in biopharmaceutical manufacturing, often reducing product yield and quality. Matriptase\u20101, a type II transmembrane serine protease, has been identified as a key contributor to unwanted proteolysis. This study investigates hepatocyte growth factor activator inhibitor\u20101 (HAI\u20101) overexpression as a strategy to mitigate matriptase\u20101\u2010mediated degradation in CHO cell culture. Using an IL\u201012 IgG1 Fc fusion protein and a multi\u2010specific antibody (M molecule) as model proteins, we employed genetic and biochemical approaches to assess the impact of Chinese hamster HAI\u20101 overexpression on protein quality and yield. Our results demonstrate that HAI\u20101 overexpression effectively inhibits matriptase\u20101 activity, achieving up to a 98% reduction in proteolytic clipping while maintaining cell growth, viability, and product quality. Compared to other protease control strategies, HAI\u20101 overexpression presents a practical and scalable solution that does not disrupt essential cellular functions. These findings establish HAI\u20101 as a key modulatory tool in CHO\u2010based protein production, with implications for reducing proteolysis of therapeutic proteins and optimizing biopharmaceutical manufacturing processes.<\/jats:p>","DOI":"10.1002\/biot.70141","type":"journal-article","created":{"date-parts":[[2025,10,27]],"date-time":"2025-10-27T06:58:58Z","timestamp":1761548338000},"source":"Crossref","is-referenced-by-count":1,"title":["Overexpression of Matriptase\u20101 Inhibitor HAI\u20101 as an Effective Strategy to Reduce Proteolytic Clipping in Chinese Hamster Ovary Cells"],"prefix":"10.1002","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-9043-8688","authenticated-orcid":false,"given":"Qinghao","family":"Zhang","sequence":"first","affiliation":[{"name":"Biologics Process Research and Development MRL Merck &amp; Co., Inc.  Rahway New Jersey USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0004-6211-2224","authenticated-orcid":false,"given":"Fnu","family":"Aapjeet","sequence":"additional","affiliation":[{"name":"Biologics Process Research and Development MRL Merck &amp; Co., Inc.  Rahway New Jersey USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0009-3935-1231","authenticated-orcid":false,"given":"Joyce","family":"Chen","sequence":"additional","affiliation":[{"name":"Biologics Process Research and Development MRL Merck &amp; Co., Inc.  Rahway New Jersey USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiaoyan","family":"Tang","sequence":"additional","affiliation":[{"name":"Biologics Process Research and Development MRL Merck &amp; Co., Inc.  Rahway New Jersey USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gregory F.","family":"Pirrone","sequence":"additional","affiliation":[{"name":"Analytical Research and Development MRL Merck &amp; Co., Inc.  Rahway New Jersey USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4866-0430","authenticated-orcid":false,"given":"Alexey A.","family":"Makarov","sequence":"additional","affiliation":[{"name":"Analytical Research and Development MRL Merck &amp; Co., Inc.  Rahway New Jersey USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Fengfei","family":"Ma","sequence":"additional","affiliation":[{"name":"Analytical Research and Development MRL Merck &amp; Co., Inc.  Rahway New Jersey USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Luke","family":"Nelson","sequence":"additional","affiliation":[{"name":"Biologics Process Research and Development MRL Merck &amp; Co., Inc.  Rahway New Jersey USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0002-3281-3869","authenticated-orcid":false,"given":"Jessica","family":"Pan","sequence":"additional","affiliation":[{"name":"Biologics Process Research and Development MRL Merck &amp; Co., Inc.  Rahway New Jersey USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1809-1441","authenticated-orcid":false,"given":"Ren","family":"Liu","sequence":"additional","affiliation":[{"name":"Biologics Process Research and Development MRL Merck &amp; Co., Inc.  Rahway New Jersey USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael J.","family":"Hohn","sequence":"additional","affiliation":[{"name":"Biologics Process Research and Development MRL Merck &amp; Co., Inc.  Rahway New Jersey USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-9258-8361","authenticated-orcid":false,"given":"David J.","family":"Busch","sequence":"additional","affiliation":[{"name":"Biologics Process Research and Development MRL Merck &amp; Co., Inc.  Rahway New Jersey USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,10,26]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biotechadv.2015.10.015"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00253\u2010024\u201013008\u20106"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1002\/bit.26731"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pgen.1010873"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1042\/bcj20200630"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.3390\/cells9041027"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1042\/BCJ20200182"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0183507"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1093\/synbio\/ysac026"},{"key":"e_1_2_10_11_1","doi-asserted-by":"crossref","unstructured":"M. D.Oberst L. Y.Chen K.Kiyomiya et\u00a0al. \u201cHAI\u20101 Regulates Activation and Expression of Matriptase a Membrane\u2010Bound Serine Protease \u201dAmerican Journal of Physiology. Cell Physiology289 no.2(2005):C462\u2013C470 https:\/\/doi.org\/10.1152\/ajpcell.00076.2005.","DOI":"10.1152\/ajpcell.00076.2005"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.2741\/2707"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.202209114"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.prp.2025.155809"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1007\/s13577\u2010024\u201001159\u20107"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jbiotec.2025.04.010"},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.1002\/2211\u20105463.12994"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.1021\/jm990412m"},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M117.815142"},{"key":"e_1_2_10_20_1","doi-asserted-by":"publisher","DOI":"10.2174\/1389203033487180"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1349\u20107006.2011.02162.x"},{"key":"e_1_2_10_22_1","doi-asserted-by":"publisher","DOI":"10.1111\/pin.12647"},{"key":"e_1_2_10_23_1","doi-asserted-by":"publisher","DOI":"10.1093\/oxfordjournals.jbchem.a022522"},{"key":"e_1_2_10_24_1","doi-asserted-by":"publisher","DOI":"10.2119\/2006\u201000022.List"},{"key":"e_1_2_10_25_1","doi-asserted-by":"publisher","DOI":"10.1242\/dev.004556"},{"key":"e_1_2_10_26_1","doi-asserted-by":"publisher","DOI":"10.2353\/ajpath.2009.090053"},{"key":"e_1_2_10_27_1","doi-asserted-by":"crossref","unstructured":"H.Xu Z.Xu I. C.Tseng et\u00a0al. \u201cMechanisms for the Control of Matriptase Activity in the Absence of Sufficient HAI\u20101 \u201dAmerican Journal of Physiology. Cell Physiology302 no.2(2012):C453\u2013C462 https:\/\/doi.org\/10.1152\/ajpcell.00344.2011.","DOI":"10.1152\/ajpcell.00344.2011"},{"key":"e_1_2_10_28_1","doi-asserted-by":"publisher","DOI":"10.1002\/bit.27016"},{"key":"e_1_2_10_29_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M117.779256"},{"key":"e_1_2_10_30_1","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M109.055640"},{"key":"e_1_2_10_31_1","doi-asserted-by":"publisher","DOI":"10.3390\/ijms17060913"},{"key":"e_1_2_10_32_1","doi-asserted-by":"publisher","DOI":"10.1290\/0411075.1"},{"key":"e_1_2_10_33_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0233866"}],"container-title":["Biotechnology Journal"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/biot.70141","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,11,30]],"date-time":"2025-11-30T07:44:16Z","timestamp":1764488656000},"score":1,"resource":{"primary":{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/10.1002\/biot.70141"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,10]]},"references-count":32,"journal-issue":{"issue":"10","published-print":{"date-parts":[[2025,10]]}},"alternative-id":["10.1002\/biot.70141"],"URL":"https:\/\/doi.org\/10.1002\/biot.70141","archive":["Portico"],"relation":{},"ISSN":["1860-6768","1860-7314"],"issn-type":[{"value":"1860-6768","type":"print"},{"value":"1860-7314","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,10]]},"article-number":"e70141"}}