{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T20:26:39Z","timestamp":1792182399951,"version":"4.4.0"},"reference-count":67,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"6801","funder":[{"DOI":"10.13039\/100032572","name":"Westlake Education Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100032572","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"The Natural Science Foundation of China","doi-asserted-by":"crossref","award":["32120103013"],"award-info":[{"award-number":["32120103013"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"crossref"}]},{"name":"National Key Research and Development Program of China","award":["2024YFC3407400"],"award-info":[{"award-number":["2024YFC3407400"]}]},{"name":"Westlake Laboratory of Life Sciences and Biomedicine"},{"name":"Pioneer and Leading Goose R&D Program of Zhejiang","award":["2024SDXHDX0002"],"award-info":[{"award-number":["2024SDXHDX0002"]}]},{"name":"Research Center for Industries of the Future (RCIF) at Westlake University"}],"content-domain":{"domain":["www.science.org"],"crossmark-restriction":true},"short-container-title":["Science"],"published-print":{"date-parts":[[2026,5,28]]},"abstract":"<jats:p>\n                    Covalent protein drugs offer therapeutic potential but are limited by slow target engagement and the absence of high-throughput selection platforms. Rapid covalent binding requires coordinated optimization of affinity, stability, and warhead geometry, which is an intrinsically multidimensional challenge. We developed a yeast display platform coupled with chemoselective modification that enables selection of fast-acting covalent proteins without increasing intrinsic warhead reactivity. Using this system, we engineered a covalent programmed death-ligand 1 (PD-L1) antagonistic nanobody with rapid cross-linking kinetics [observed rate constant (\n                    <jats:italic toggle=\"yes\">k<\/jats:italic>\n                    <jats:sub>obs<\/jats:sub>\n                    ) = 0.18 min\n                    <jats:sup>\u22121<\/jats:sup>\n                    , half-life (\n                    <jats:italic toggle=\"yes\">t<\/jats:italic>\n                    <jats:sub>1\/2<\/jats:sub>\n                    ) = 3.8 min] and improved tumor suppression compared with envafolimab and atezolizumab. Similarly, we engineered a fast-acting covalent interleukin-18 (\n                    <jats:italic toggle=\"yes\">k<\/jats:italic>\n                    <jats:sub>obs<\/jats:sub>\n                    = 0.54 min\n                    <jats:sup>\u22121<\/jats:sup>\n                    ,\n                    <jats:italic toggle=\"yes\">t<\/jats:italic>\n                    <jats:sub>1\/2<\/jats:sub>\n                    = 1.3 min) and a covalent miniprotein targeting the receptor binding domain of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), demonstrating applicability across protein modalities.\n                  <\/jats:p>","DOI":"10.1126\/science.adv3081","type":"journal-article","created":{"date-parts":[[2026,4,2]],"date-time":"2026-04-02T17:59:27Z","timestamp":1775152767000},"update-policy":"https:\/\/doi.org\/10.34133\/aaas_crossmark","source":"Crossref","is-referenced-by-count":5,"title":["A high-throughput selection system for fast-acting covalent protein drugs"],"prefix":"10.1126","volume":"392","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-8498-5007","authenticated-orcid":true,"given":"Qiongxuan","family":"Fan","sequence":"first","affiliation":[{"name":"College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, China."},{"name":"Research Center for Industries of the Future and Key Laboratory of Structural Biology of Zhejiang Province, School of Life Sciences, Westlake University, Hangzhou, Zhejiang, China."},{"name":"Westlake Laboratory of Life Sciences and Biomedicine, Hangzhou, Zhejiang, China."},{"name":"Institute of Biology, Westlake Institute for Advanced Study, Hangzhou, Zhejiang, China."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6365-8243","authenticated-orcid":true,"given":"Jiahao","family":"Mei","sequence":"additional","affiliation":[{"name":"College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, China."},{"name":"Research Center for Industries of the Future and Key Laboratory of Structural Biology of Zhejiang Province, School of Life Sciences, Westlake University, Hangzhou, Zhejiang, China."},{"name":"Westlake Laboratory of Life Sciences and Biomedicine, Hangzhou, Zhejiang, China."},{"name":"Institute of Biology, Westlake Institute for Advanced Study, Hangzhou, Zhejiang, China."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-9137-9785","authenticated-orcid":true,"given":"Tian","family":"Li","sequence":"additional","affiliation":[{"name":"Research Center for Industries of the Future and Key Laboratory of Structural Biology of Zhejiang Province, School of Life Sciences, Westlake University, Hangzhou, Zhejiang, China."},{"name":"Westlake Laboratory of Life Sciences and Biomedicine, Hangzhou, Zhejiang, China."},{"name":"Institute of Biology, Westlake Institute for Advanced Study, Hangzhou, Zhejiang, China."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0006-7434-0771","authenticated-orcid":true,"given":"Chuanlong","family":"Zang","sequence":"additional","affiliation":[{"name":"Research Center for Industries of the Future and Key Laboratory of Structural Biology of Zhejiang Province, School of Life Sciences, Westlake University, Hangzhou, Zhejiang, China."},{"name":"Westlake Laboratory of Life Sciences and Biomedicine, Hangzhou, Zhejiang, China."},{"name":"Institute of Biology, Westlake Institute for Advanced Study, Hangzhou, Zhejiang, China."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-1178-4474","authenticated-orcid":true,"given":"Mengjiao","family":"Li","sequence":"additional","affiliation":[{"name":"Research Center for Industries of the Future and Key Laboratory of Structural Biology of Zhejiang Province, School of Life Sciences, Westlake University, Hangzhou, Zhejiang, China."},{"name":"Westlake Laboratory of Life Sciences and Biomedicine, Hangzhou, Zhejiang, China."},{"name":"Institute of Biology, Westlake Institute for Advanced Study, Hangzhou, Zhejiang, China."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0008-6966-7491","authenticated-orcid":true,"given":"Jing","family":"Tang","sequence":"additional","affiliation":[{"name":"Westlake Laboratory of Life Sciences and Biomedicine, Hangzhou, Zhejiang, China."},{"name":"School of Medicine, Westlake University, Hangzhou, Zhejiang, China."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6129-1492","authenticated-orcid":true,"given":"You","family":"Xu","sequence":"additional","affiliation":[{"name":"Research Center for Industries of the Future and Key Laboratory of Structural Biology of Zhejiang Province, School of Life Sciences, Westlake University, Hangzhou, Zhejiang, China."},{"name":"Westlake Laboratory of Life Sciences and Biomedicine, Hangzhou, Zhejiang, China."},{"name":"Institute of Biology, Westlake Institute for Advanced Study, Hangzhou, Zhejiang, China."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-5565-3951","authenticated-orcid":true,"given":"Ge","family":"Yu","sequence":"additional","affiliation":[{"name":"Research Center for Industries of the Future and Key Laboratory of Structural Biology of Zhejiang Province, School of Life Sciences, Westlake University, Hangzhou, Zhejiang, China."},{"name":"Westlake Laboratory of Life Sciences and Biomedicine, Hangzhou, Zhejiang, China."},{"name":"Institute of Biology, Westlake Institute for Advanced Study, Hangzhou, Zhejiang, China."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dandan","family":"Liu","sequence":"additional","affiliation":[{"name":"Zhejiang Provincial Key Laboratory for Cancer Molecular Cell Biology, Life Sciences Institute, Zhejiang University, Hangzhou, Zhejiang, China."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0206-3502","authenticated-orcid":true,"given":"Kai","family":"Chen","sequence":"additional","affiliation":[{"name":"Research Center for Industries of the Future and Key Laboratory of Structural Biology of Zhejiang Province, School of Life Sciences, Westlake University, Hangzhou, Zhejiang, China."},{"name":"Westlake Laboratory of Life Sciences and Biomedicine, Hangzhou, Zhejiang, China."},{"name":"Institute of Biology, Westlake Institute for Advanced Study, Hangzhou, Zhejiang, China."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6739-2932","authenticated-orcid":true,"given":"Bing","family":"Yang","sequence":"additional","affiliation":[{"name":"Zhejiang Provincial Key Laboratory for Cancer Molecular Cell Biology, Life Sciences Institute, Zhejiang University, Hangzhou, Zhejiang, China."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9639-2907","authenticated-orcid":true,"given":"Jing","family":"Huang","sequence":"additional","affiliation":[{"name":"Research Center for Industries of the Future and Key Laboratory of Structural Biology of Zhejiang Province, School of Life Sciences, Westlake University, Hangzhou, Zhejiang, China."},{"name":"Westlake Laboratory of Life Sciences and Biomedicine, Hangzhou, Zhejiang, China."},{"name":"Institute of Biology, Westlake Institute for Advanced Study, Hangzhou, Zhejiang, China."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8783-8380","authenticated-orcid":true,"given":"Ting","family":"Zhou","sequence":"additional","affiliation":[{"name":"Westlake Laboratory of Life Sciences and Biomedicine, Hangzhou, Zhejiang, China."},{"name":"School of Medicine, Westlake University, Hangzhou, Zhejiang, China."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1962-8677","authenticated-orcid":true,"given":"Bobo","family":"Dang","sequence":"additional","affiliation":[{"name":"Research Center for Industries of the Future and Key Laboratory of Structural Biology of Zhejiang Province, School of Life Sciences, Westlake University, Hangzhou, Zhejiang, China."},{"name":"Westlake Laboratory of Life Sciences and Biomedicine, Hangzhou, Zhejiang, China."},{"name":"Institute of Biology, Westlake Institute for Advanced Study, Hangzhou, Zhejiang, China."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"221","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41573-022-00542-z"},{"key":"e_1_3_2_3_2","doi-asserted-by":"publisher","DOI":"10.1039\/C9CS00720B"},{"key":"e_1_3_2_4_2","doi-asserted-by":"publisher","DOI":"10.1002\/cpz1.419"},{"key":"e_1_3_2_5_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.drudis.2015.05.005"},{"key":"e_1_3_2_6_2","doi-asserted-by":"publisher","DOI":"10.1161\/01.CIR.101.10.1206"},{"key":"e_1_3_2_7_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2020.05.028"},{"key":"e_1_3_2_8_2","doi-asserted-by":"publisher","DOI":"10.1021\/jacs.1c08521"},{"key":"e_1_3_2_9_2","doi-asserted-by":"publisher","DOI":"10.1021\/jacs.1c11554"},{"key":"e_1_3_2_10_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.cbpa.2021.102106"},{"key":"e_1_3_2_11_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chempr.2022.07.012"},{"key":"e_1_3_2_12_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2023.09.004"},{"key":"e_1_3_2_13_2","doi-asserted-by":"publisher","DOI":"10.1039\/D3SC01921G"},{"key":"e_1_3_2_14_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibs.2019.12.008"},{"key":"e_1_3_2_15_2","doi-asserted-by":"publisher","DOI":"10.3389\/fonc.2020.01182"},{"key":"e_1_3_2_16_2","doi-asserted-by":"publisher","DOI":"10.1038\/nri2747"},{"key":"e_1_3_2_17_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.copbio.2011.06.012"},{"key":"e_1_3_2_18_2","doi-asserted-by":"publisher","DOI":"10.1002\/anie.202412843"},{"key":"e_1_3_2_19_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.accounts.6b00321"},{"key":"e_1_3_2_20_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1521020113"},{"key":"e_1_3_2_21_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-025-09286-3"},{"key":"e_1_3_2_22_2","volume-title":"Lehninger Principles of Biochemistry","author":"Nelson D. L.","year":"2021","unstructured":"D. L. Nelson, M. M. Cox, Lehninger Principles of Biochemistry (W. H. Freeman, ed. 8, 2021).","edition":"8"},{"key":"e_1_3_2_23_2","doi-asserted-by":"publisher","DOI":"10.1038\/nbt0697-553"},{"key":"e_1_3_2_24_2","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4939-2748-7_8"},{"key":"e_1_3_2_25_2","doi-asserted-by":"publisher","DOI":"10.1038\/celldisc.2017.4"},{"key":"e_1_3_2_26_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2115001118"},{"key":"e_1_3_2_27_2","doi-asserted-by":"publisher","DOI":"10.1002\/onco.13817"},{"key":"e_1_3_2_28_2","doi-asserted-by":"publisher","DOI":"10.1038\/s42003-020-0916-2"},{"key":"e_1_3_2_29_2","doi-asserted-by":"publisher","DOI":"10.1038\/nature14404"},{"key":"e_1_3_2_30_2","doi-asserted-by":"publisher","DOI":"10.1158\/2326-6066.CIR-15-0077-T"},{"key":"e_1_3_2_31_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJMoa1200694"},{"key":"e_1_3_2_32_2","doi-asserted-by":"publisher","DOI":"10.2174\/0113816128248929230920071937"},{"key":"e_1_3_2_33_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.it.2012.07.006"},{"key":"e_1_3_2_34_2","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.181.3.1627"},{"key":"e_1_3_2_35_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.semcancer.2022.05.002"},{"key":"e_1_3_2_36_2","doi-asserted-by":"publisher","DOI":"10.1158\/1078-0432.CCR-07-4740"},{"key":"e_1_3_2_37_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-020-2422-6"},{"key":"e_1_3_2_38_2","doi-asserted-by":"publisher","DOI":"10.7717\/peerj.13626"},{"key":"e_1_3_2_39_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0809086106"},{"key":"e_1_3_2_40_2","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms6340"},{"key":"e_1_3_2_41_2","unstructured":"A. Ring T. Zhou S. Fischer Interleukin-18 variants and methods of use. US Patent Application 2019\/0070262 A1 (2019)."},{"key":"e_1_3_2_42_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.abd9909"},{"key":"e_1_3_2_43_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41587-020-0733-7"},{"key":"e_1_3_2_44_2","doi-asserted-by":"publisher","DOI":"10.1039\/D1CC01773J"},{"key":"e_1_3_2_45_2","doi-asserted-by":"publisher","DOI":"10.1021\/jacs.3c04833"},{"key":"e_1_3_2_46_2","doi-asserted-by":"publisher","DOI":"10.1021\/jacs.4c15843"},{"key":"e_1_3_2_47_2","doi-asserted-by":"publisher","DOI":"10.3389\/fimmu.2024.1490911"},{"key":"e_1_3_2_48_2","doi-asserted-by":"publisher","DOI":"10.1021\/acscentsci.3c00288"},{"key":"e_1_3_2_49_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41589-021-00770-1"},{"key":"e_1_3_2_50_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-022-05235-6"},{"key":"e_1_3_2_51_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.4193"},{"key":"e_1_3_2_52_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.4169"},{"key":"e_1_3_2_53_2","doi-asserted-by":"publisher","DOI":"10.1002\/jcc.20084"},{"key":"e_1_3_2_54_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444909052925"},{"key":"e_1_3_2_55_2","doi-asserted-by":"publisher","DOI":"10.1107\/S0907444904019158"},{"key":"e_1_3_2_56_2","doi-asserted-by":"publisher","DOI":"10.1002\/wcms.70019"},{"key":"e_1_3_2_57_2","doi-asserted-by":"publisher","DOI":"10.1002\/jcc.21287"},{"key":"e_1_3_2_58_2","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.4067"},{"key":"e_1_3_2_59_2","doi-asserted-by":"publisher","DOI":"10.1002\/jcc.21367"},{"key":"e_1_3_2_60_2","doi-asserted-by":"publisher","DOI":"10.1021\/ci3003649"},{"key":"e_1_3_2_61_2","doi-asserted-by":"publisher","DOI":"10.1021\/ci300363c"},{"key":"e_1_3_2_62_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jpcb.3c06662"},{"key":"e_1_3_2_63_2","doi-asserted-by":"publisher","DOI":"10.1002\/jcc.21787"},{"key":"e_1_3_2_64_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1101046108"},{"key":"e_1_3_2_65_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41422-022-00656-4"},{"key":"e_1_3_2_66_2","unstructured":"Y. Xu The MD simulation trajectories of IB101\/PDL1 and KN035_L108C_6H\/PDL1 systems Zenodo (2026); 10.5281\/zenodo.18417074"},{"key":"e_1_3_2_67_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.orglett.7b02522"},{"key":"e_1_3_2_68_2","doi-asserted-by":"publisher","DOI":"10.1021\/acs.jcim.0c01301"}],"container-title":["Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.science.org\/doi\/pdf\/10.1126\/science.adv3081","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,5,28]],"date-time":"2026-05-28T18:00:51Z","timestamp":1779991251000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.science.org\/doi\/10.1126\/science.adv3081"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,5,28]]},"references-count":67,"journal-issue":{"issue":"6801","published-print":{"date-parts":[[2026,5,28]]}},"alternative-id":["10.1126\/science.adv3081"],"URL":"https:\/\/doi.org\/10.1126\/science.adv3081","relation":{},"ISSN":["0036-8075","1095-9203"],"issn-type":[{"value":"0036-8075","type":"print"},{"value":"1095-9203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,5,28]]},"assertion":[{"value":"2024-12-13","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-12-03","order":1,"name":"revised","label":"Revised","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-03-10","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-05-28","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"eadv3081"}}