{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,26]],"date-time":"2026-02-26T15:14:48Z","timestamp":1772118888646,"version":"3.50.1"},"reference-count":46,"publisher":"Wiley","issue":"14","license":[{"start":{"date-parts":[[2025,7,18]],"date-time":"2025-07-18T00:00:00Z","timestamp":1752796800000},"content-version":"vor","delay-in-days":17,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Proteomics"],"published-print":{"date-parts":[[2025,7]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>\n                    SOBERANA 02 is a safe and effective anti\u2010SARS\u2010CoV\u20102 conjugate vaccine, produced using the maleimide\u2010thiol chemistry. In this vaccine, Cys\n                    <jats:sup>538<\/jats:sup>\n                    in the recombinant receptor binding domain (RBD) of SARS\u2010CoV\u20102 is linked through a\n                    <jats:italic>N<\/jats:italic>\n                    \u2010thiosuccinimidopropionyl linker to lysine residues of tetanus toxoid (TT) preparation. LC\u2010MS\/MS analysis revealed the complex protein composition of TT. The fifteen most abundant proteins account for approximately 78% of the total protein mass. The detoxified tetanus neurotoxin (d\u2010TeNT) was the most abundant protein (30%\u201356%) regardless of the quantification method used. LC\u2010MS\/MS analysis of d\u2010TeNT activation reaction with BMPS (3\u2010maleimidopropionic acid\n                    <jats:italic>N<\/jats:italic>\n                    \u2010hydroxysuccinimide ester) showed that 102 (95%) of the 107 lysine residues incorporated a maleimide group. Of them, only 22 Lys residues (20%) were cross\u2010linked to the RBD\n                    <jats:italic>C<\/jats:italic>\n                    \u2010terminal tryptic peptide (\n                    <jats:sup>538<\/jats:sup>\n                    CVNF\n                    <jats:sup>541<\/jats:sup>\n                    \u2010HHHHHH), probably due to steric hindrance. Affinity chromatography prior to LC\u2010MS\/MS analysis was critical to identify the conjugation sites. Linear peptides carrying a conjugated lysine residue and type 2 peptides with stabilized linker forms (hydrolyzed and transcyclized), allowed the identification of twelve and eighteen conjugation sites, respectively. The RBD was also conjugated, but to a lesser extent, to ten other low\u2010abundance carrier proteins. This study represents the first report of a conjugation site assignment in a TT\u2010based conjugate vaccine.\n                  <\/jats:p>","DOI":"10.1002\/pmic.13978","type":"journal-article","created":{"date-parts":[[2025,7,18]],"date-time":"2025-07-18T10:20:53Z","timestamp":1752834053000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["LC\u2010MS\/MS Characterization of SOBERANA 02, a Receptor Binding Domain\u2010Tetanus Toxoid Conjugate Vaccine Against SARS\u2010CoV\u20102"],"prefix":"10.1002","volume":"25","author":[{"given":"Olivia","family":"Mart\u00ednez","sequence":"first","affiliation":[{"name":"Finlay Institute of Vaccines  Havana Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Darielys","family":"Santana\u2010Medero","sequence":"additional","affiliation":[{"name":"Finlay Institute of Vaccines  Havana Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Satomy","family":"Pousa","sequence":"additional","affiliation":[{"name":"Center for Genetic Engineering and Biotechnology  Havana Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jean Pierre","family":"Soubal","sequence":"additional","affiliation":[{"name":"Finlay Institute of Vaccines  Havana Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Arielis","family":"Rodr\u00edguez\u2010Ulloa","sequence":"additional","affiliation":[{"name":"Center for Genetic Engineering and Biotechnology  Havana Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pablo E.","family":"Ramos\u2010Berm\u00fadez","sequence":"additional","affiliation":[{"name":"Center for Genetic Engineering and Biotechnology  Havana Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vladimir","family":"Besada","sequence":"additional","affiliation":[{"name":"Center for Genetic Engineering and Biotechnology  Havana Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Raine","family":"Garrido","sequence":"additional","affiliation":[{"name":"Finlay Institute of Vaccines  Havana Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Paulo","family":"Carvalho","sequence":"additional","affiliation":[{"name":"Carlos Chagas Institute\/FioCruz Rua Professor Algacyr Munhoz Mader  Curitiba Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michel","family":"Batista","sequence":"additional","affiliation":[{"name":"Carlos Chagas Institute\/FioCruz Rua Professor Algacyr Munhoz Mader  Curitiba Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Katharina","family":"Zetl","sequence":"additional","affiliation":[{"name":"Department Proteomics and Signal Transduction Max Planck Institute of Biochemistry  Martinsried Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jacek R.","family":"Wi\u015bniewski","sequence":"additional","affiliation":[{"name":"Department Proteomics and Signal Transduction Max Planck Institute of Biochemistry  Martinsried Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tamy","family":"Boggiano","sequence":"additional","affiliation":[{"name":"Center for Molecular Immunology  Havana Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yury","family":"Vald\u00e9s\u2010Balb\u00edn","sequence":"additional","affiliation":[{"name":"Finlay Institute of Vaccines  Havana Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dagmar","family":"Garc\u00eda\u2010Rivera","sequence":"additional","affiliation":[{"name":"Finlay Institute of Vaccines  Havana Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Daniel G.","family":"Rivera","sequence":"additional","affiliation":[{"name":"Finlay Institute of Vaccines  Havana Cuba"},{"name":"Laboratory of Synthetic and Biomolecular Chemistry Faculty of Chemistry University of Havana  La Habana Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vicente","family":"Verez\u2010Bencomo","sequence":"additional","affiliation":[{"name":"Finlay Institute of Vaccines  Havana Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8875-3642","authenticated-orcid":false,"given":"Luis Javier","family":"Gonz\u00e1lez","sequence":"additional","affiliation":[{"name":"Center for Genetic Engineering and Biotechnology  Havana Cuba"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2025,7,18]]},"reference":[{"key":"e_1_2_9_2_1","doi-asserted-by":"publisher","DOI":"10.3390\/bioengineering9020072"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.coi.2021.07.003"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1021\/acscentsci.1c00216"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1021\/acschembio.1c00272"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.3389\/fbioe.2023.1287551"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2022.05.082"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1039\/d1cb00200g"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.lana.2022.100423"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.lana.2024.100750"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.nbt.2022.08.002"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1177\/1469066719893492"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00216\u2010021\u201003721\u2010w"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.aca.2016.11.073"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.chembiol.2013.01.010"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.analchem.1c04025.s001"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00216\u2010021\u201003569\u20100"},{"key":"e_1_2_9_18_1","doi-asserted-by":"publisher","DOI":"10.3390\/pathogens9060513"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.microc.2024.111025"},{"key":"e_1_2_9_20_1","doi-asserted-by":"publisher","DOI":"10.1002\/rcm.9859"},{"key":"e_1_2_9_21_1","doi-asserted-by":"publisher","DOI":"10.1039\/d4an01449a"},{"key":"e_1_2_9_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/S1044\u20100305(03)00327\u20101"},{"key":"e_1_2_9_23_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41467\u2010019\u201011337\u2010z"},{"key":"e_1_2_9_24_1","doi-asserted-by":"publisher","DOI":"10.1021\/pr501321h"},{"key":"e_1_2_9_25_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jsb.2010.10.014"},{"key":"e_1_2_9_26_1","doi-asserted-by":"publisher","DOI":"10.1002\/rcm.9660"},{"key":"e_1_2_9_27_1","doi-asserted-by":"publisher","DOI":"10.1002\/chem.202003951"},{"key":"e_1_2_9_28_1","doi-asserted-by":"publisher","DOI":"10.1021\/bc5005262"},{"key":"e_1_2_9_29_1","doi-asserted-by":"publisher","DOI":"10.3390\/proteomes7020015"},{"key":"e_1_2_9_30_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jprot.2014.07.012"},{"key":"e_1_2_9_31_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature10098"},{"key":"e_1_2_9_32_1","doi-asserted-by":"publisher","DOI":"10.1074\/mcp.m500230\u2010mcp200"},{"key":"e_1_2_9_33_1","doi-asserted-by":"publisher","DOI":"10.1186\/1471\u20102105\u20108\u20104"},{"key":"e_1_2_9_34_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jchromb.2017.04.009"},{"key":"e_1_2_9_35_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.vaccine.2006.12.033"},{"key":"e_1_2_9_36_1","doi-asserted-by":"publisher","DOI":"10.1016\/0003\u20109861(58)90014\u20106"},{"key":"e_1_2_9_37_1","doi-asserted-by":"publisher","DOI":"10.1021\/ac504689z"},{"key":"e_1_2_9_38_1","doi-asserted-by":"publisher","DOI":"10.1074\/mcp.m113.031591"},{"key":"e_1_2_9_39_1","doi-asserted-by":"publisher","DOI":"10.1074\/mcp.m111.010587"},{"key":"e_1_2_9_40_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth1113"},{"key":"e_1_2_9_41_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.analchem.3c04682"},{"key":"e_1_2_9_42_1","doi-asserted-by":"publisher","DOI":"10.1074\/mcp.M115.049296"},{"key":"e_1_2_9_43_1","doi-asserted-by":"publisher","DOI":"10.1021\/ac049169b"},{"key":"e_1_2_9_44_1","doi-asserted-by":"publisher","DOI":"10.1021\/acs.analchem.1c01317.s001"},{"key":"e_1_2_9_45_1","doi-asserted-by":"publisher","DOI":"10.1007\/s13361\u2010018\u20101917\u2010y"},{"key":"e_1_2_9_46_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkac1040"},{"key":"e_1_2_9_47_1","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gkae1011"}],"container-title":["PROTEOMICS"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/pmic.13978","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,21]],"date-time":"2025-10-21T17:53:36Z","timestamp":1761069216000},"score":1,"resource":{"primary":{"URL":"https:\/\/analyticalsciencejournals.onlinelibrary.wiley.com\/doi\/10.1002\/pmic.13978"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,7]]},"references-count":46,"journal-issue":{"issue":"14","published-print":{"date-parts":[[2025,7]]}},"alternative-id":["10.1002\/pmic.13978"],"URL":"https:\/\/doi.org\/10.1002\/pmic.13978","archive":["Portico"],"relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/2024.12.27.630511","asserted-by":"object"}]},"ISSN":["1615-9853","1615-9861"],"issn-type":[{"value":"1615-9853","type":"print"},{"value":"1615-9861","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,7]]},"assertion":[{"value":"2025-03-05","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-05-27","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2025-07-18","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e13978"}}