A Bead‐Based Screening Platform for Identifying Monoclonal Antibodies That Disrupt PD‐1/PD‐L1 Interactions
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Le résumé fourni par la source
Monoclonal antibodies (mAbs) targeting immune checkpoint pathways such as programmed cell death protein 1 (PD‐1)/PD‐L1 are central to modern immunotherapy, yet scalable methods to assess their functional blockade remain limited. We present a bead‐based flow cytometry assay for quantifying the inhibition of PD‐1/PD‐L1 interaction by antibodies. Recombinant human PD‐1 protein was conjugated to polystyrene beads, and its interaction with recombinant human PD‐L1 protein labeled with a fluorochrome was measured. The inhibitory activity of an anti‐PD‐L1 mAb was quantified based on their ability to disrupt this interaction. The assay was validated for intra‐ and inter‐assay precision, in addition, functionality was confirmed using a T cell coculture assay. The assay demonstrated dose‐dependent inhibition by the αPD‐L1 mAb, with a calculated mean IC 50 of 3.122 µg/mL. The method proved to be reproducible for the determination of antibody blocking activity, with relative standard deviation (RSD) < 20% between three independent runs. At the concentration approximating the IC 50 detected on the bead assay, the antibody significantly restored CD69 expression on the T cell surface ( p = 0.0001) in a coculture in vitro system. In addition, the methodology could successfully distinguish the blocking capacity of two anti‐PD‐L1 antibodies with different affinities. This high‐throughput compatible platform offers a reliable tool for screening PD‐1/PD‐L1 blocking antibodies, supporting immunotherapy discovery and development.
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Le contrôle bibliographique ouvert
DOI retrouvé dans Crossref DOI retrouvé ; titre concordant.
- Titre Crossref
- A Bead‐Based Screening Platform for Identifying Monoclonal Antibodies That Disrupt PD‐1/PD‐L1 Interactions
- Date Crossref
- 01/01/2026
- Éditeur
- Wiley
- Type
- journal-article
Ce recoupement confirme des métadonnées liées au DOI. Il ne confirme ni la méthode ni les conclusions de l’étude, et il ne compte pas comme une seconde source scientifique indépendante.
Où se fait cette recherche
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Universidade Federal do Rio Grande do Sul pays non établi dans la noticeUniversité ou école supérieure
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Universidade Federal de Ciências da Saúde de Porto Alegre pays non établi dans la noticeUniversité ou école supérieure
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Universidade de São Paulo pays non établi dans la noticeUniversité ou école supérieure
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Hospital Universitário da Universidade de São Paulo pays non établi dans la noticeÉtablissement de santé
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Federal University of Health Sciences of Porto Alegre Immunotherapy Laboratory pays non établi dans la noticeUniversité ou école supérieure
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University of São Paulo Department of Microbiology pays non établi dans la noticeUniversité ou école supérieure
Universidade Federal do Rio Grande do Sul, Universidade Federal de Ciências da Saúde de Porto Alegre et Universidade de São Paulo, avec 3 autres affiliations.
Une affiliation ne permet pas de déduire la nationalité d’un auteur.