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Lookup NU author(s): Alba Méndez Alejandre, Dr Ben RaymondORCiD, Professor Matthias TrostORCiD, Dr Jose Luis Marin-RubioORCiD
This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).
Copyright © 2023 Méndez-Alejandre, Raymond, Trost and Marín-Rubio. Phagosome acidification and proteolysis are essential processes in the immune response to contain and eliminate pathogens. In recent years, there has been an increased desire for a rapid and accurate method of assessing these processes in real-time. Here, we outline the development of a multiplexed assay that allows simultaneous monitoring of phagosome acidification and proteolysis in the same sample using silica beads conjugated to pHrodo and DQ BSA. We describe in detail how to prepare the bi-functional particles and show proof of concept using differentially activated macrophages. This multiplexed spectrophotometric assay allows rapid and accurate assessment of phagosome acidification and proteolysis in real-time and could provide valuable information for understanding the immune response to pathogen invasion.
Author(s): Mendez-Alejandre A, Raymond BBA, Trost M, Marin-Rubio JL
Publication type: Article
Publication status: Published
Journal: Frontiers in Immunology
Year: 2023
Volume: 14
Online publication date: 10/08/2023
Acceptance date: 17/07/2023
Date deposited: 15/09/2023
ISSN (electronic): 1664-3224
Publisher: Frontiers Media SA
URL: https://doi.org/10.3389/fimmu.2023.1204223
DOI: 10.3389/fimmu.2023.1204223
Data Access Statement: The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.
PubMed id: 37638042
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