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To elucidate the molecular interactions governing the formation of bovine lactoferrin (BLF)\u2212\u03ba-carrageenan (\u03baCG) bionanostructures, the thermodynamic and kinetic parameters of BLF\u2212\u03baCG binding were determined using Steady-state fluorescence (FS) and surface plasmon resonance (SPR) spectroscopies. Formation of a 1:1 BLF-\u03baCG complex is thermodynamically favorable at equilibrium (\u221247.00 \u2264 \u0394GFSo \u2264 \u221246.15 kJ mol\u20131) and is predominantly enthalpy-driven (\u0394HFSo = \u221257.43 kJ mol\u20131). Kinetic analysis by SPR revealed that the interaction proceeds through a transition complex ([BLF \u2013 \u03baCG]\u2021), with complex formation occurring more rapidly through association of the free molecules (32.53 \u2264 \u0394Ga\u2021 \u2264 38.52 kJ mol\u20131) than through dissociation of the thermodynamically stable complex (77.61 \u2264 \u0394Gd\u2021 \u2264 81.62 kJ mol\u20131). Consistent with these findings, the energetic barrier for dissociation (Ed\u2013298.2\u00a0K\u2021 = 96.46 kJ mol\u20131) was higher than for association (Ea\u2013298.2\u00a0K\u2021 = 42.88 kJ mol\u20131). Increasing the ionic strength altered the BLF\u2212\u03baCG interaction mechanism, converting the binding process from a multistep pathway to a single-step process.<\/jats:p>","DOI":"10.1021\/acsomega.6c08383","type":"journal-article","created":{"date-parts":[[2026,9,26]],"date-time":"2026-09-26T15:01:20Z","timestamp":1790434880000},"page":"60360-60370","source":"Crossref","is-referenced-by-count":0,"title":["Lactoferrin\/\u03ba-Carrageenan\nInteractions: Thermodynamic\nand Kinetic Insights through Surface Plasmon Resonance and Steady-State\nFluorescence"],"prefix":"10.1021","volume":"11","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9221-0421","authenticated-orcid":true,"given":"Yara Luiza","family":"Coelho Zampier","sequence":"first","affiliation":[{"name":"Federal University of Vic\u0327osa , , PH Rolfs Avenue , ,","place":["Vic\u0327osa-MG, Brazil, 36570-000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6376-4812","authenticated-orcid":true,"given":"Isabela A.","family":"Marques","sequence":"additional","affiliation":[{"name":"Federal University of Vic\u0327osa , , PH Rolfs Avenue , ,","place":["Vic\u0327osa-MG, Brazil, 36570-000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2991-8117","authenticated-orcid":true,"given":"A\u0301lvaro Javier","family":"Patin\u0303o-Agudelo","sequence":"additional","affiliation":[{"name":"Federal University of Vic\u0327osa , , PH Rolfs Avenue , ,","place":["Vic\u0327osa-MG, Brazil, 36570-000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hauster Maximiler C.","family":"de Paula","sequence":"additional","affiliation":[{"name":"Federal University of Vic\u0327osa , , PH Rolfs Avenue , ,","place":["Vic\u0327osa-MG, Brazil, 36570-000"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eliara A.","family":"Hudson","sequence":"additional","affiliation":[{"name":"Federal University of Vic\u0327osa , , Av. P. H. Rolfs s\/s , ,","place":["Vic\u0327osa-MG, Brazil, 36570900"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana Clarissa S.","family":"Pires","sequence":"additional","affiliation":[{"name":"Federal University of Vic\u0327osa , , Av. P. H. 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