Optimized Methods for the Surface Immobilization of Collagens and Collagen Binding Assays

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dc.contributor.authorChaher, Nadia
dc.contributor.authorDigilio, Giuseppe
dc.contributor.authorLacerda, Sara
dc.contributor.authorBotnar, Rene M.
dc.contributor.authorPhinikaridou, Alkystis
dc.date.accessioned2025-01-20T20:15:04Z
dc.date.available2025-01-20T20:15:04Z
dc.date.issued2023
dc.description.abstractFibrosis occurs in various tissues as a reparative response to injury or damage. If excessive, however, fibrosis can lead to tissue scarring and organ failure, which is associated with high morbidity and mortality. Collagen is a key driver of fibrosis, with type I and type III collagen being the primary types involved in many fibrotic diseases. Unlike conventional protocols used to immobilize other proteins (e.g., elastin, albumin, fibronectin, etc.), comprehensive protocols to reproducibly immobilize different types of collagens in order to produce stable coatings are not readily available. Immobilizing collagen is surprisingly challenging because multiple experimental conditions may affect the efficiency of immobilization, including the type of collagen, the pH, the temperature, and the type of microplate used. Here, a detailed protocol to reproducibly immobilize and quantify type I and III collagens resulting in stable and reproducible gels/films is provided. Furthermore, this work demonstrates how to perform, analyze, and interpret in vitro time-resolved fluorescence binding studies to investigate the interactions between collagens and candidate collagen-binding compounds (e.g., a peptide conjugated to a metal chelate carrying, for example, europium [Eu(III)]). Such an approach can be universally applied to various biomedical applications, including the field of molecular imaging to develop targeted imaging probes, drug development, cell toxicity studies, cell proliferation studies, and immunoassays.
dc.fuente.origenWOS
dc.identifier.doi10.3791/64720
dc.identifier.issn1940-087X
dc.identifier.urihttps://doi.org/10.3791/64720
dc.identifier.urihttps://repositorio.uc.cl/handle/11534/92238
dc.identifier.wosidWOS:000971111100028
dc.issue.numero193
dc.language.isoen
dc.revistaJove-journal of visualized experiments
dc.rightsacceso restringido
dc.subject.ods03 Good Health and Well-being
dc.subject.odspa03 Salud y bienestar
dc.titleOptimized Methods for the Surface Immobilization of Collagens and Collagen Binding Assays
dc.typeartículo
sipa.indexWOS
sipa.trazabilidadWOS;2025-01-12
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