Isolation of the B Cell Immune Synapse for Proteomic Analysis

Diogo M. Cunha; Sara Hernández-Pérez; Pieta K. Mattila

doi:10.1007/978-1-0716-3135-5_25

Isolation of the B Cell Immune Synapse for Proteomic Analysis

Diogo M. Cunha, Sara Hernández-Pérez, Pieta K. Mattila^*

^*Corresponding author for this work

Research output: Chapter in Book/Conference proceeding › Chapter › Scientific › peer-review

1 Citation (Scopus)

Abstract

Recent technical developments have fueled increasing utilization of proteomics to gain new insights into various aspects of cellular behavior. In this chapter, we describe a method to specifically isolate immune synapses from mouse primary B cells. The method utilizes antibody-coated magnetic beads to induce the formation of the immune synapses and describes a protocol for the extraction of the cell-bead adhesions for mass spectrometry analysis. Finally, this method enables unveiling the large-scale protein content of the immune synapse.

Original language	English
Title of host publication	Methods in Molecular Biology
Publisher	Humana Press Inc.
Pages	393-408
Number of pages	16
Volume	2654
ISBN (Print)	978-1-0716-3134-8
DOIs	https://doi.org/10.1007/978-1-0716-3135-5_25
Publication status	Published - 28 Apr 2023
MoE publication type	A3 Part of a book or another research book

Publication series

Name	Methods in Molecular Biology
Volume	2654
ISSN (Print)	1064-3745
ISSN (Electronic)	1940-6029

Funding

We thank Dr. Alexandre Carisey (St Jude’s Research Hospital, Memphis, USA) for his generous help with setting up the protocol. We also acknowledge Prof. Jordan Orange and Dr. Anna Meyer for their pioneering efforts in the immune synapse isolation in NK cells (Department of Pediatrics, Vagelos College of Physicians and Surgeons, Columbia University Irving Medical Center, New York, New York, USA). Turku Proteomics Facility of Turku Bioscience and Biocenter Finland are acknowledged for providing the research infrastructures. This work was supported by the Academy of Finland (decision number: 339810 to PKM), Sigrid Juselius foundation (to PKM), Erasmus+ program (to DMC), the Finnish Cultural Foundation (Suomen Kulttuurirahasto; to SHP), and by InFLAMES Flagship Programme of the Academy of Finland (deci-sion number: 337530).

Funders	Funder number
Columbia University Irving Medical Center, New York, New York
Turku Proteomics Facility of Turku Bioscience
Department of Pediatrics, University of Florida
Vagelos College of Physicians and Surgeons, Columbia University
Academy of Finland	339810
Finnish Cultural Foundation	337530
Sigrid Jusélius Foundation
Biocenter Finland

Keywords

B cells
Cell adhesion
Immunological synapse immune synapse
Mass spectrometry
Proteomics

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10.1007/978-1-0716-3135-5_25

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abstract = "Recent technical developments have fueled increasing utilization of proteomics to gain new insights into various aspects of cellular behavior. In this chapter, we describe a method to specifically isolate immune synapses from mouse primary B cells. The method utilizes antibody-coated magnetic beads to induce the formation of the immune synapses and describes a protocol for the extraction of the cell-bead adhesions for mass spectrometry analysis. Finally, this method enables unveiling the large-scale protein content of the immune synapse.",

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AB - Recent technical developments have fueled increasing utilization of proteomics to gain new insights into various aspects of cellular behavior. In this chapter, we describe a method to specifically isolate immune synapses from mouse primary B cells. The method utilizes antibody-coated magnetic beads to induce the formation of the immune synapses and describes a protocol for the extraction of the cell-bead adhesions for mass spectrometry analysis. Finally, this method enables unveiling the large-scale protein content of the immune synapse.

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ER -

Isolation of the B Cell Immune Synapse for Proteomic Analysis

Abstract

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Keywords

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