Titre
A stepwise protocol to coat aAPC beads prevents out-competition of anti-CD3 mAb and consequent experimental artefacts.
Type
article
Institution
UNIL/CHUV/Unisanté + institutions partenaires
Périodique
Auteur(s)
Fuertes Marraco, S.A.
Auteure/Auteur
Baumgaertner, P.
Auteure/Auteur
Legat, A.
Auteure/Auteur
Rufer, N.
Auteure/Auteur
Speiser, D.E.
Auteure/Auteur
Liens vers les personnes
Liens vers les unités
ISSN
1872-7905
Statut éditorial
Publié
Date de publication
2012
Volume
385
Numéro
1-2
Première page
90
Dernière page/numéro d’article
95
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov'tPublication Status: ppublish
Résumé
Artificial antigen-presenting cells (aAPC) are widely used for both clinical and basic research applications, as cell-based or bead-based scaffolds, combining immune synapse components of interest. Adequate and controlled preparation of aAPCs is crucial for subsequent immunoassays. We reveal that certain proteins such as activatory anti-CD3 antibody can be out-competed by other proteins (e.g. inhibitory receptor ligands such as PDL1:Fc) during the coating of aAPC beads, under the usually performed coating procedures. This may be misleading, as we found that decreased CD8 T cell activity was not due to inhibitory receptor triggering but rather because of unexpectedly low anti-CD3 antibody density on the beads upon co-incubation with inhibitory receptor ligands. We propose an optimized protocol, and emphasize the need to quality-control the coating of proteins on aAPC beads prior to their use in immunoassays.
PID Serval
serval:BIB_101C0CB4CDE2
PMID
Date de création
2012-11-22T13:27:57.196Z
Date de création dans IRIS
2025-05-20T15:49:19Z