{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,21]],"date-time":"2026-04-21T07:40:58Z","timestamp":1776757258376,"version":"3.51.2"},"reference-count":28,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2018,11,23]],"date-time":"2018-11-23T00:00:00Z","timestamp":1542931200000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/www.springer.com\/tdm"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Nat Protoc"],"published-print":{"date-parts":[[2019,1]]},"DOI":"10.1038\/s41596-018-0083-9","type":"journal-article","created":{"date-parts":[[2018,11,22]],"date-time":"2018-11-22T16:18:17Z","timestamp":1542903497000},"page":"86-99","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":65,"title":["Efficient and irreversible antibody\u2013cysteine bioconjugation using carbonylacrylic reagents"],"prefix":"10.1038","volume":"14","author":[{"given":"Barbara","family":"Bernardim","sequence":"first","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3470-8299","authenticated-orcid":false,"given":"Maria J.","family":"Matos","sequence":"additional","affiliation":[]},{"given":"Xhenti","family":"Ferhati","sequence":"additional","affiliation":[]},{"given":"Ismael","family":"Compa\u00f1\u00f3n","sequence":"additional","affiliation":[]},{"given":"Ana","family":"Guerreiro","sequence":"additional","affiliation":[]},{"given":"Padma","family":"Akkapeddi","sequence":"additional","affiliation":[]},{"given":"Antonio C. B.","family":"Burtoloso","sequence":"additional","affiliation":[]},{"given":"Gonzalo","family":"Jim\u00e9nez-Os\u00e9s","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5597-8127","authenticated-orcid":false,"given":"Francisco","family":"Corzana","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6594-8917","authenticated-orcid":false,"given":"Gon\u00e7alo J. L.","family":"Bernardes","sequence":"additional","affiliation":[]}],"member":"297","published-online":{"date-parts":[[2018,11,23]]},"reference":[{"key":"83_CR1","doi-asserted-by":"publisher","first-page":"114","DOI":"10.1038\/nchem.2415","volume":"8","author":"V Chudasama","year":"2016","unstructured":"Chudasama, V., Maruani, A. & Caddick, S. Recent advances in the construction of antibody\u2013drug conjugates. Nat. Chem. 8, 114\u2013119 (2016).","journal-title":"Nat. Chem."},{"key":"83_CR2","doi-asserted-by":"publisher","first-page":"917","DOI":"10.1038\/nchembio.1959","volume":"11","author":"L Xue","year":"2015","unstructured":"Xue, L., Karpenko, I. A., Hiblot, J. & Johnsson, K. Imaging and manipulating proteins in live cells through covalent labeling. Nat. Chem. Biol. 11, 917\u2013923 (2015).","journal-title":"Nat. Chem. Biol."},{"key":"83_CR3","doi-asserted-by":"publisher","first-page":"1088","DOI":"10.1038\/nchem.2685","volume":"8","author":"T Rodrigues","year":"2016","unstructured":"Rodrigues, T. & Bernardes, G. J. L. The missing link. Nat. Chem. 8, 1088\u20131090 (2016).","journal-title":"Nat. Chem."},{"key":"83_CR4","doi-asserted-by":"publisher","first-page":"103","DOI":"10.1038\/nchem.2393","volume":"8","author":"N Krall","year":"2015","unstructured":"Krall, N., da Cruz, F. P., Boutureira, O. & Bernardes, G. J. L. Site-selective protein-modification chemistry for basic biology and drug development. Nat. Chem. 8, 103\u2013113 (2015).","journal-title":"Nat. Chem."},{"key":"83_CR5","doi-asserted-by":"publisher","first-page":"326","DOI":"10.1038\/nchembio.1792","volume":"11","author":"JI MacDonald","year":"2015","unstructured":"MacDonald, J. I., Munch, H. K., Moore, T. & Francis, M. B. One-step site-specific modification of native proteins with 2-pyridinecarboxyaldehydes. Nat. Chem. Biol. 11, 326\u2013331 (2015).","journal-title":"Nat. Chem. Biol."},{"key":"83_CR6","doi-asserted-by":"publisher","first-page":"687","DOI":"10.1038\/nature15739","volume":"526","author":"EV Vinogradova","year":"2015","unstructured":"Vinogradova, E. V., Zhang, C., Spokoyny, A. M., Pentelute, B. L. & Buchwald, S. L. Organometallic palladium reagents for cysteine bioconjugation. Nature 526, 687\u2013691 (2015).","journal-title":"Nature"},{"key":"83_CR7","doi-asserted-by":"publisher","first-page":"2174","DOI":"10.1021\/cr500399p","volume":"115","author":"O Boutureira","year":"2015","unstructured":"Boutureira, O. & Bernardes, G. J. L. Advances in chemical protein modification. Chem. Rev. 115, 2174\u20132195 (2015).","journal-title":"Chem. Rev."},{"key":"83_CR8","doi-asserted-by":"publisher","first-page":"4004","DOI":"10.1021\/jacs.7b12874","volume":"140","author":"MJ Matos","year":"2018","unstructured":"Matos, M. J. et al. Chemo- and regioselective lysine modification on native proteins. J. Am. Chem. Soc. 140, 4004\u20134017 (2018).","journal-title":"J. Am. Chem. Soc."},{"key":"83_CR9","doi-asserted-by":"publisher","first-page":"315","DOI":"10.1038\/nrd.2016.268","volume":"16","author":"A Beck","year":"2017","unstructured":"Beck, A., Goetsch, L., Dumontet, C. & Corva\u00efa, N. Strategies and challenges for the next generation of antibody\u2013drug conjugates. Nat. Rev. Drug Discov. 16, 315\u2013337 (2017).","journal-title":"Nat. Rev. Drug Discov."},{"key":"83_CR10","doi-asserted-by":"publisher","first-page":"15","DOI":"10.1146\/annurev-med-050311-201823","volume":"64","author":"EL Sievers","year":"2013","unstructured":"Sievers, E. L. & Senter, P. D. Antibody-drug conjugates in cancer therapy. Ann. Rev. Med. 64, 15\u201329 (2013).","journal-title":"Ann. Rev. Med."},{"key":"83_CR11","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms13128","volume":"7","author":"B Bernardim","year":"2016","unstructured":"Bernardim, B. et al. Stoichiometric and irreversible cysteine-selective protein modification using carbonylacrylic reagents. Nat. Commun. 7, 13128 (2016).","journal-title":"Nat. Commun."},{"key":"83_CR12","doi-asserted-by":"publisher","first-page":"925","DOI":"10.1038\/nbt.1480","volume":"26","author":"JR Junutula","year":"2008","unstructured":"Junutula, J. R. et al. Site-specific conjugation of a cytotoxic drug to an antibody improves the therapeutic index. Nat. Biotechnol. 26, 925\u2013932 (2008).","journal-title":"Nat. Biotechnol."},{"key":"83_CR13","doi-asserted-by":"publisher","first-page":"4185","DOI":"10.1039\/C8SC00256H","volume":"9","author":"BJ Stenton","year":"2018","unstructured":"Stenton, B. J., Oliveira, B. L., Matos, M. J., Sinatra, L. & Bernardes, G. J. L. A thioether-directed palladium-cleavable linker for targeted bioorthogonal drug decaging. Chem. Sci. 9, 4185\u20134189 (2018).","journal-title":"Chem. Sci."},{"key":"83_CR14","doi-asserted-by":"publisher","first-page":"697","DOI":"10.1038\/nchembio.2416","volume":"13","author":"CB Rosen","year":"2017","unstructured":"Rosen, C. B. & Francis, M. B. Targeting the N terminus for site-selective protein modification. Nat. Chem. Biol. 13, 697\u2013705 (2017).","journal-title":"Nat. Chem. Biol."},{"key":"83_CR15","doi-asserted-by":"publisher","first-page":"630","DOI":"10.1002\/asia.200800427","volume":"4","author":"JM Chalker","year":"2009","unstructured":"Chalker, J. M., Bernardes, G. J. L., Lin, Y. A. & Davis, B. G. Chemical modification of proteins at cysteine: opportunities in chemistry and biology. Chem. Asian J. 4, 630\u2013640 (2009).","journal-title":"Chem. Asian J."},{"key":"83_CR16","doi-asserted-by":"crossref","unstructured":"Pimenta G\u00f3is, P. M., Ravasco, J., Faustino, H. & Trindade, A. Bioconjugation with maleimides: a useful tool for chemical biology. Chem. Eur. J. doi:10.1002\/chem.201803174 (2018).","DOI":"10.1002\/chem.201803174"},{"key":"83_CR17","doi-asserted-by":"publisher","first-page":"1432","DOI":"10.1002\/anie.201508118","volume":"55","author":"D Kalia","year":"2015","unstructured":"Kalia, D., Malekar Pushpa, V. & Parthasarathy, M. Exocyclic olefinic maleimides: synthesis and application for stable and thiol\u2010selective bioconjugation. Angew. Chem. Int. Ed. Engl. 55, 1432\u20131435 (2015).","journal-title":"Angew. Chem. Int. Ed. Engl."},{"key":"83_CR18","doi-asserted-by":"publisher","first-page":"1059","DOI":"10.1038\/nbt.2968","volume":"32","author":"RP Lyon","year":"2014","unstructured":"Lyon, R. P. et al. Self-hydrolyzing maleimides improve the stability and pharmacological properties of antibody-drug conjugates. Nat. Biotechnol. 32, 1059\u20131062 (2014).","journal-title":"Nat. Biotechnol."},{"key":"83_CR19","doi-asserted-by":"publisher","first-page":"2433","DOI":"10.1096\/fj.10-180331","volume":"25","author":"I Vaneycken","year":"2011","unstructured":"Vaneycken, I. et al. Preclinical screening of anti-HER2 nanobodies for molecular imaging of breast cancer. FASEB J. 25, 2433\u20132446 (2011).","journal-title":"FASEB J."},{"key":"83_CR20","doi-asserted-by":"publisher","first-page":"733","DOI":"10.1038\/nbt.3212","volume":"33","author":"RP Lyon","year":"2015","unstructured":"Lyon, R. P. et al. Reducing hydrophobicity of homogeneous antibody-drug conjugates improves pharmacokinetics and therapeutic index. Nat. Biotechnol. 33, 733\u2013735 (2015).","journal-title":"Nat. Biotechnol."},{"key":"83_CR21","doi-asserted-by":"publisher","first-page":"18365","DOI":"10.1021\/jacs.7b10702","volume":"139","author":"AM Freedy","year":"2017","unstructured":"Freedy, A. M. et al. Chemoselective installation of amine bonds on proteins through aza-Michael ligation. J. Am. Chem. Soc. 139, 18365\u201318375 (2017).","journal-title":"J. Am. Chem. Soc."},{"key":"83_CR22","doi-asserted-by":"publisher","first-page":"733","DOI":"10.1002\/anie.201608292","volume":"56","author":"MR Levengood","year":"2017","unstructured":"Levengood, M. R. et al. Orthogonal cysteine protection enables homogeneous multi-drug antibody\u2013drug conjugates. Angew. Chem. Int. Ed. Engl. 56, 733\u2013737 (2017).","journal-title":"Angew. Chem. Int. Ed. Engl."},{"key":"83_CR23","doi-asserted-by":"publisher","first-page":"631","DOI":"10.1038\/nbt.2289","volume":"30","author":"PD Senter","year":"2012","unstructured":"Senter, P. D. & Sievers, E. L. The discovery and development of brentuximab vedotin for use in relapsed Hodgkin lymphoma and systemic anaplastic large cell lymphoma. Nat. Biotechnol. 30, 631\u2013637 (2012).","journal-title":"Nat. Biotechnol."},{"key":"83_CR24","doi-asserted-by":"publisher","first-page":"479","DOI":"10.1021\/jm00137a001","volume":"24","author":"PL Carl","year":"1981","unstructured":"Carl, P. L., Chakravarty, P. K. & Katzenellenbogen, J. A. A novel connector linkage applicable in prodrug design. J. Med. Chem. 24, 479\u2013480 (1981).","journal-title":"J. Med. Chem."},{"key":"83_CR25","doi-asserted-by":"publisher","first-page":"6342","DOI":"10.1021\/jm2007613","volume":"54","author":"JJ Cui","year":"2011","unstructured":"Cui, J. J. et al. Structure based drug design of crizotinib (PF-02341066), a potent and selective dual inhibitor of mesenchymal\u2013epithelial transition factor (c-MET) kinase and anaplastic lymphoma kinase (ALK). J. Med. Chem. 54, 6342\u20136363 (2011).","journal-title":"J. Med. Chem."},{"key":"83_CR26","doi-asserted-by":"publisher","first-page":"3200","DOI":"10.1158\/1078-0432.CCR-05-2804","volume":"12","author":"SS Brack","year":"2006","unstructured":"Brack, S. S., Silacci, M., Birchler, M. & Neri, D. Tumor-targeting properties of novel antibodies specific to the large isoform of tenascin-c. Clin. Cancer Res. 12, 3200\u20133208 (2006).","journal-title":"Clin. Cancer Res."},{"key":"83_CR27","doi-asserted-by":"publisher","first-page":"397","DOI":"10.2967\/jnumed.112.111310","volume":"54","author":"DA Heuveling","year":"2013","unstructured":"Heuveling, D. A. et al. Phase 0 microdosing PET study using the human mini antibody F16SIP in head and neck cancer patients. J. Nucl. Med. 54, 397\u2013401 (2013).","journal-title":"J. Nucl. Med."},{"key":"83_CR28","doi-asserted-by":"publisher","first-page":"1670","DOI":"10.1002\/ijc.30569","volume":"140","author":"R G\u00e9bleux","year":"2016","unstructured":"G\u00e9bleux, R., Stringhini, M., Casanova, R., Soltermann, A. & Neri, D. Non\u2010internalizing antibody\u2013drug conjugates display potent anti\u2010cancer activity upon proteolytic release of monomethyl auristatin E in the subendothelial extracellular matrix. Int. J. Cancer 140, 1670\u20131679 (2016).","journal-title":"Int. J. Cancer"}],"container-title":["Nature Protocols"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/www.nature.com\/articles\/s41596-018-0083-9","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"http:\/\/www.nature.com\/articles\/s41596-018-0083-9.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"http:\/\/www.nature.com\/articles\/s41596-018-0083-9.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,5,20]],"date-time":"2023-05-20T22:27:58Z","timestamp":1684621678000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41596-018-0083-9"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,11,23]]},"references-count":28,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2019,1]]}},"alternative-id":["83"],"URL":"https:\/\/doi.org\/10.1038\/s41596-018-0083-9","relation":{},"ISSN":["1754-2189","1750-2799"],"issn-type":[{"value":"1754-2189","type":"print"},{"value":"1750-2799","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,11,23]]},"assertion":[{"value":"23 November 2018","order":1,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}]}}