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^*

^*Tämän työn vastaava kirjoittaja

Tutkimustuotos: Artikkeli kirjassa/raportissa/konferenssijulkaisussa › Luku › Tieteellinen › vertaisarvioitu

1 Sitaatiot (Scopus)

Abstrakti

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.

Alkuperäiskieli	Englanti
Otsikko	Methods in Molecular Biology
Kustantaja	Humana Press Inc.
Sivut	393-408
Sivumäärä	16
Vuosikerta	2654
ISBN (painettu)	978-1-0716-3134-8
DOI - pysyväislinkit	https://doi.org/10.1007/978-1-0716-3135-5_25
Tila	Julkaistu - 28 huhtik. 2023
OKM-julkaisutyyppi	A3 Kirjan osa tai toinen tutkimuskirja

Julkaisusarja

Nimi	Methods in Molecular Biology
Vuosikerta	2654
ISSN (painettu)	1064-3745
ISSN (elektroninen)	1940-6029

Rahoitus

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).

Rahoittajat	Rahoittajan numero
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
Suomen Kulttuurirahasto	337530
Sigrid Juséliuksen säätiö
Biocenter Finland

Pääsy asiakirjaan

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

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Viittausmuodot

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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.",

keywords = "B cells, Cell adhesion, Immunological synapse immune synapse, Mass spectrometry, Proteomics",

author = "Cunha, {Diogo M.} and Sara Hern{\'a}ndez-P{\'e}rez and Mattila, {Pieta K.}",

note = "Publisher Copyright: {\textcopyright} 2023, The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.",

year = "2023",

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AU - Cunha, Diogo M.

AU - Hernández-Pérez, Sara

AU - Mattila, Pieta K.

PY - 2023/4/28

Y1 - 2023/4/28

N2 - 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.

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.

KW - B cells

KW - Cell adhesion

KW - Immunological synapse immune synapse

KW - Mass spectrometry

KW - Proteomics

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U2 - 10.1007/978-1-0716-3135-5_25

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

M3 - Chapter

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AN - SCOPUS:85158146981

SN - 978-1-0716-3134-8

VL - 2654

T3 - Methods in Molecular Biology

SP - 393

EP - 408

BT - Methods in Molecular Biology

PB - Humana Press Inc.

ER -

Isolation of the B Cell Immune Synapse for Proteomic Analysis

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