A stepwise protocol to coat aAPC beads prevents out-competition of anti-CD3 mAb and consequent experimental artefacts.

Détails

ID Serval
serval:BIB_101C0CB4CDE2
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Institution
Titre
A stepwise protocol to coat aAPC beads prevents out-competition of anti-CD3 mAb and consequent experimental artefacts.
Périodique
Journal of Immunological Methods
Auteur⸱e⸱s
Fuertes Marraco S.A., Baumgaertner P., Legat A., Rufer N., Speiser D.E.
ISSN
1872-7905 (Electronic)
ISSN-L
0022-1759
Statut éditorial
Publié
Date de publication
2012
Volume
385
Numéro
1-2
Pages
90-95
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.
Pubmed
Web of science
Création de la notice
22/11/2012 15:27
Dernière modification de la notice
20/08/2019 13:36
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