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Chem."],"published-print":{"date-parts":[[2023,11]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Bispecific T cell engagers (BiTEs), a subset of bispecific antibodies (bsAbs), can promote a targeted cancer cell\u2019s death by bringing it close to a cytotoxic T cell. Checkpoint inhibitory T cell engagers (CiTEs) comprise a BiTE core with an added immunomodulatory protein, which serves to reverse cancer-cell immune-dampening strategies, improving efficacy. So far, protein engineering has been the main approach to generate bsAbs and CiTEs, but improved chemical methods for their generation have recently been developed. Homogeneous fragment-based bsAbs constructed from fragment antigen-binding regions (Fabs) can be generated using click chemistry. Here we describe a chemical method to generate biotin-functionalized three-protein conjugates, which include two CiTE molecules, one containing an anti-PD-1 Fab and the other containing an immunomodulatory enzyme,\n                    <jats:italic>Salmonella typhimurium<\/jats:italic>\n                    sialidase. The CiTEs\u2019 efficacy was shown to be superior to that of the simpler BiTE scaffold, with the sialidase-containing CiTE inducing substantially enhanced T cell-mediated cytotoxicity in vitro. The chemical method described here, more generally, enables the generation of multi-protein constructs with further biological applications.\n                  <\/jats:p>","DOI":"10.1038\/s41557-023-01280-4","type":"journal-article","created":{"date-parts":[[2023,7,24]],"date-time":"2023-07-24T12:01:51Z","timestamp":1690200111000},"page":"1636-1647","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":43,"title":["Chemical generation of checkpoint inhibitory T cell engagers for the treatment of cancer"],"prefix":"10.1038","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2944-2676","authenticated-orcid":false,"given":"Peter A.","family":"Szijj","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Melissa A.","family":"Gray","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6816-8326","authenticated-orcid":false,"given":"Mikaela K.","family":"Ribi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Calise","family":"Bahou","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5992-5900","authenticated-orcid":false,"given":"Jo\u00e3o C. F.","family":"Nogueira","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4482-2754","authenticated-orcid":false,"given":"Carolyn R.","family":"Bertozzi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8876-3285","authenticated-orcid":false,"given":"Vijay","family":"Chudasama","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2023,7,24]]},"reference":[{"key":"1280_CR1","doi-asserted-by":"publisher","first-page":"945","DOI":"10.1080\/14712598.2022.2040987","volume":"22","author":"N Gera","year":"2022","unstructured":"Gera, N. The evolution of bispecific antibodies. Expert Opin. Biol. Ther. 22, 945\u2013949 (2022).","journal-title":"Expert Opin. Biol. Ther."},{"key":"1280_CR2","doi-asserted-by":"publisher","first-page":"1055","DOI":"10.1016\/S1470-2045(22)00335-7","volume":"23","author":"LE Budde","year":"2022","unstructured":"Budde, L. E. et al. Safety and efficacy of mosunetuzumab, a bispecific antibody, in patients with relapsed or refractory follicular lymphoma: a single-arm, multicentre, phase 2 study. Lancet Oncol. 23, 1055\u20131065 (2022).","journal-title":"Lancet Oncol."},{"key":"1280_CR3","doi-asserted-by":"publisher","first-page":"131","DOI":"10.1007\/s12272-023-01433-6","volume":"46","author":"Y Hong","year":"2023","unstructured":"Hong, Y., Nam, S. M. & Moon, A. Antibody\u2013drug conjugates and bispecific antibodies targeting cancers: applications of click chemistry. Arch. Pharm. Res. 46, 131\u2013148 (2023).","journal-title":"Arch. Pharm. 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The renaissance of chemically generated bispecific antibodies. Nat. Rev. Chem. 5, 78\u201392 (2021).","journal-title":"Nat. Rev. Chem."},{"key":"1280_CR7","doi-asserted-by":"publisher","first-page":"14404","DOI":"10.1021\/jacs.2c00129","volume":"144","author":"RJ Taylor","year":"2022","unstructured":"Taylor, R. J., Geeson, M. B., Journeaux, T. & Bernardes, G. J. L. Chemical and enzymatic methods for post-translational protein-protein conjugation. J. Am. Chem. Soc. 144, 14404\u201314419 (2022).","journal-title":"J. Am. Chem. Soc."},{"key":"1280_CR8","doi-asserted-by":"publisher","first-page":"e77319","DOI":"10.1371\/journal.pone.0077319","volume":"8","author":"A Moura","year":"2013","unstructured":"Moura, A., Savageau, M. A. & Alves, R. Relative amino acid composition signatures of organisms and environments. 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Tuning the hydrolytic stability of next generation maleimide cross-linkers enables access to albumin-antibody fragment conjugates and tri-scFvs. Bioconjug. Chem. 29, 486\u2013492 (2018).","journal-title":"Bioconjug. Chem."},{"key":"1280_CR12","doi-asserted-by":"publisher","first-page":"5490","DOI":"10.1016\/j.bmcl.2017.09.065","volume":"27","author":"JT Patterson","year":"2017","unstructured":"Patterson, J. T. et al. PSMA-targeted bispecific Fab conjugates that engage T cells. Bioorg. Med. Chem. Lett. 27, 5490\u20135495 (2017).","journal-title":"Bioorg. Med. Chem. Lett."},{"key":"1280_CR13","doi-asserted-by":"publisher","first-page":"3647","DOI":"10.1016\/j.bmcl.2017.07.021","volume":"27","author":"JT Patterson","year":"2017","unstructured":"Patterson, J. T. et al. Chemically generated IgG2 bispecific antibodies through disulfide bridging. Bioorg. Med. Chem. Lett. 27, 3647\u20133652 (2017).","journal-title":"Bioorg. Med. Chem. 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A bioorthogonal click chemistry toolbox for targeted synthesis of branched and well-defined protein-protein conjugates. Angew. Chem. Int. Ed. 59, 12885\u201312893 (2020).","journal-title":"Angew. Chem. Int. Ed."},{"key":"1280_CR27","doi-asserted-by":"publisher","first-page":"41","DOI":"10.3390\/antib8030041","volume":"8","author":"WR Strohl","year":"2019","unstructured":"Strohl, W. R. & Naso, M. Bispecific T-cell redirection versus chimeric antigen receptor (CAR)-T cells as approaches to kill cancer cells. Antibodies 8, 41 (2019).","journal-title":"Antibodies"},{"key":"1280_CR28","doi-asserted-by":"publisher","first-page":"348","DOI":"10.1158\/2326-6066.CIR-18-0622","volume":"7","author":"D Neri","year":"2019","unstructured":"Neri, D. Antibody-cytokine fusions: versatile products for the modulation of anticancer immunity. Cancer Immunol. Res. 7, 348\u2013354 (2019).","journal-title":"Cancer Immunol. 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Programmable half-life and anti-tumour effects of bispecific T-cell engager-albumin fusions with tuned FcRn affinity. Commun. Biol. 4, 310 (2021).","journal-title":"Commun. Biol."},{"key":"1280_CR34","doi-asserted-by":"publisher","first-page":"3739","DOI":"10.1021\/acs.molpharmaceut.7b00497","volume":"14","author":"L Liu","year":"2017","unstructured":"Liu, L. et al. Albumin binding domain fusing R\/K-X-X-R\/K sequence for enhancing tumor delivery of doxorubicin. Mol. Pharm. 14, 3739\u20133749 (2017).","journal-title":"Mol. Pharm."}],"container-title":["Nature Chemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41557-023-01280-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41557-023-01280-4","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41557-023-01280-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,11,3]],"date-time":"2023-11-03T09:07:38Z","timestamp":1699002458000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41557-023-01280-4"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,7,24]]},"references-count":34,"journal-issue":{"issue":"11","published-print":{"date-parts":[[2023,11]]}},"alternative-id":["1280"],"URL":"https:\/\/doi.org\/10.1038\/s41557-023-01280-4","relation":{"has-preprint":[{"id-type":"doi","id":"10.26434\/chemrxiv-2022-5p97s-v3","asserted-by":"object"}]},"ISSN":["1755-4330","1755-4349"],"issn-type":[{"value":"1755-4330","type":"print"},{"value":"1755-4349","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,7,24]]},"assertion":[{"value":"11 October 2022","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"21 June 2023","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"24 July 2023","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"M.A.G. and C.R.B. are inventors of a patent filed by Stanford University (international publication no. WO2018006034A1) titled \u2018Conjugates for targeted cell-surface editing\u2019 published on 4 January 2018 and licensed by Palleon Pharmaceuticals on 27 June 2017. C.R.B. is a cofounder and Scientific Advisory Board member of Palleon Pharmaceuticals, Enable Bioscience, Redwood Biosciences (a subsidiary of Catalent), InterVenn Biosciences, Lycia Therapeutics, Grace Science LLC and OliLux Biosciences. V.C. is a director of the spin-out ThioLogics and is an inventor of the patent filed by UCL Business PLC (European patent EP-2464654-B1) titled \u2018Thiol protecting group\u2019 published on 8 October 2014, and other patents derived thereof, as well as directly related patents in other jurisdictions that fall within the patent family. The remaining authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}]}}