{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T20:58:09Z","timestamp":1788382689115,"version":"build-2803163510"},"reference-count":29,"publisher":"Wiley","license":[{"start":{"date-parts":[[2026,2,8]],"date-time":"2026-02-08T00:00:00Z","timestamp":1770508800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc\/4.0\/"},{"start":{"date-parts":[[2026,2,8]],"date-time":"2026-02-08T00:00:00Z","timestamp":1770508800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100007129","name":"Natural Science Foundation of Shandong Province","doi-asserted-by":"publisher","award":["ZR2024QC068"],"award-info":[{"award-number":["ZR2024QC068"]}],"id":[{"id":"10.13039\/501100007129","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Plant Biotechnology Journal"],"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    Efficient purification remains a key technical challenge affecting the recovery efficiency of recombinant proteins in plant\u2010based systems. Complex metabolites, particularly polyphenols, often cause recombinant protein aggregation during purification. In this study, we identified two key polyphenol oxidase genes,\n                    <jats:italic>PPOa<\/jats:italic>\n                    and\n                    <jats:italic>PPOb<\/jats:italic>\n                    from\n                    <jats:italic>Nicotiana benthamiana<\/jats:italic>\n                    , as responsible for these effects. Using CRISPR\u2010Cas9, we generated two\n                    <jats:italic>ppoa;ppob<\/jats:italic>\n                    double knockout lines that significantly improved the purification of virus surface proteins like SARS\u2010CoV\u20102 Spike trimer and influenza HA trimer. These lines showed reduced polyphenol\u2013protein interactions, minimised aggregation, and higher purification yields. Our work establishes a clean, high\u2010efficiency\n                    <jats:italic>N. benthamiana<\/jats:italic>\n                    chassis for scalable recombinant protein production.\n                  <\/jats:p>","DOI":"10.1111\/pbi.70575","type":"journal-article","created":{"date-parts":[[2026,2,9]],"date-time":"2026-02-09T05:23:21Z","timestamp":1770614601000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Knocking Out Two Polyphenol Oxidase Genes Significantly Improves Recombinant Protein Purification in\n                    <i>Nicotiana benthamiana<\/i>"],"prefix":"10.1111","author":[{"given":"Hai\u2010Ping","family":"Diao","sequence":"first","affiliation":[{"name":"Qingdao Municipal Key Laboratory of Plant Molecular Pharming Tobacco Research Institute, Chinese Academy of Agricultural Sciences  Qingdao China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hui\u2010Xin","family":"Meng","sequence":"additional","affiliation":[{"name":"Qingdao Municipal Key Laboratory of Plant Molecular Pharming Tobacco Research Institute, Chinese Academy of Agricultural Sciences  Qingdao China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xue\u2010Jiao","family":"Xu","sequence":"additional","affiliation":[{"name":"Qingdao Municipal Key Laboratory of Plant Molecular Pharming Tobacco Research Institute, Chinese Academy of Agricultural Sciences  Qingdao China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zeng\u2010Lin","family":"Zhang","sequence":"additional","affiliation":[{"name":"Qingdao Municipal Key Laboratory of Plant Molecular Pharming Tobacco Research Institute, Chinese Academy of Agricultural Sciences  Qingdao China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jian\u2010Feng","family":"Zhang","sequence":"additional","affiliation":[{"name":"Beijing Life Science Academy (BLSA)  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yong\u2010Feng","family":"Guo","sequence":"additional","affiliation":[{"name":"Qingdao Municipal Key Laboratory of Plant Molecular Pharming Tobacco Research Institute, Chinese Academy of Agricultural Sciences  Qingdao China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Inhwan","family":"Hwang","sequence":"additional","affiliation":[{"name":"Department of Life Science Pohang University of Science and Technology  Pohang Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3474-2542","authenticated-orcid":false,"given":"Shi\u2010Jian","family":"Song","sequence":"additional","affiliation":[{"name":"Qingdao Municipal Key Laboratory of Plant Molecular Pharming Tobacco Research Institute, Chinese Academy of Agricultural Sciences  Qingdao China"},{"name":"Beijing Life Science Academy (BLSA)  Beijing China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2026,2,8]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.3389\/fpls.2018.01893"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41577-025-01207-9"},{"key":"e_1_2_9_4_1","doi-asserted-by":"crossref","DOI":"10.1016\/j.xplc.2025.101676","article-title":"Plant\u2010Derived Vaccines: Advances in Delivery Approaches","volume":"7","author":"Diao H.\u2010P.","year":"2025","journal-title":"Plant 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