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Bi-functional particles for real-time phagosome acidification and proteolysis multiplex assay in macrophages

Lookup NU author(s): Alba Méndez Alejandre, Dr Ben RaymondORCiD, Professor Matthias TrostORCiD, Dr Jose Luis Marin-RubioORCiD

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This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).

Abstract

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.

Publication metadata

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

PubMed id: 37638042

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