{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,20]],"date-time":"2026-07-20T13:04:18Z","timestamp":1784552658546,"version":"3.55.0"},"reference-count":47,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2026,6,2]],"date-time":"2026-06-02T00:00:00Z","timestamp":1780358400000},"content-version":"vor","delay-in-days":1,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"},{"start":{"date-parts":[[2026,6,1]],"date-time":"2026-06-01T00:00:00Z","timestamp":1780272000000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Biotechnology Journal"],"published-print":{"date-parts":[[2026,6]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    O\u2010xylosylation of glycine\u2013serine (GS) linkers in multispecific antibodies or fusion proteins introduces product heterogeneity, posing critical challenges for biomanufacturing quality control and elevating potential immunogenicity risks. This post\u2010translational modification is primarily catalyzed by xylosyltransferase 2 (\n                    <jats:italic>Xylt2<\/jats:italic>\n                    ) in chinese hamster ovary (CHO) cells. To address this, we generated\n                    <jats:italic>Xylt2<\/jats:italic>\n                    \u2010deficient CHO cells via both clustered regularly interspaced short palindromic repeats (CRISPR)\/CRISPR\u2010associated protein 9 (Cas9) and zinc finger nuclease (ZFN)\u2010mediated gene knockout, which resulted in the complete elimination of O\u2010xylosylation as verified by mass spectrometry in a GS linker\u2010containing bispecific antibody. Furthermore, because heparan sulfate proteoglycans (HSPGs), key cell\u2010surface receptors for ligand binding and internalization, rely on O\u2010xylosylation for their proper biosynthesis and function, the\n                    <jats:italic>Xylt2<\/jats:italic>\n                    knockout also enhanced the titer of human bone morphogenetic protein 2 (hBMP2), which undergoes HSPG\u2010dependent cellular uptake, with up to a 2.5\u2010fold increase from 9.62 to 34.09\u00a0\u00b5g\/mL in engineered cells compared to wild\u2010type CHO controls. Collectively, our results demonstrate that\n                    <jats:italic>Xylt2<\/jats:italic>\n                    \u2010deficient cell lines provide a genetic approach to produce recombinant proteins without O\u2010xylosylation, as well as for enhancing the titer of hBMP2.\n                  <\/jats:p>","DOI":"10.1002\/biot.70256","type":"journal-article","created":{"date-parts":[[2026,6,2]],"date-time":"2026-06-02T07:08:46Z","timestamp":1780384126000},"source":"Crossref","is-referenced-by-count":0,"title":["<i>Xylt2<\/i>\n                    Knockout Eliminates O\u2010Xylosylation and Enhances Bone Morphogenetic Protein 2 Production in Chinese Hamster Ovary Cells"],"prefix":"10.1002","volume":"21","author":[{"given":"Lin","family":"Han","sequence":"first","affiliation":[{"name":"Cell Line Development WuXi Biologics  Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nana","family":"Wu","sequence":"additional","affiliation":[{"name":"Cell Line 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