Isolation and Purification of B Cells from Human Peripheral Blood

Published: July 31, 2023

Abstract

Source: Tzeng, S. J. The Isolation, Differentiation, and Quantification of Human Antibody-secreting B Cells from Blood: ELISpot as a Functional Readout of Humoral Immunity. J. Vis. Exp. (2016)

In this video, we demonstrate the isolation of B cells from human peripheral blood using biotinylated antibodies specific for non-B cells and streptavidin-conjugated magnetic microbeads.

Protocol

All procedures involving human participants have been performed in compliance with the institutional, national, and international guidelines for human welfare and have been reviewed by the local institutional review board 1. Isolation and Purification of Human Peripheral Blood B Cells Draw ~ 10 mL of blood from the median cubital vein (in the cubital fossa anterior to the elbow) into a 15-mL tube containing K2EDTA (1.5 to 2.0 mg/mL blood) and immediately…

Divulgaciones

The authors have nothing to disclose.

Materials

BD Vacutainer K2E BD Biosciences 367525 10 ml tube
Ficoll-Paque Plus GE Healthcare 17-1440-02 Endotoxin-free
Trypan blue 0.5% solution Biological Industries 03-102-1B Dye
IMag Human B lymphocyte enrichment set BD Biosciences 558007 Reagent
Streptavidin magnetic microbeads BD Biosciences 9000810 Reagent
15 ml Falcon tubes BD Falcon 352196 Plastic ware
Biotinylated CD27 mAb Biolegend 302804 Clone O323
Biotinylated anti-human CD27 Biolegend 302804 Clone O323
Biotinylated anti-human CD27 eBioscience 13-0279-80 Clone O323

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Isolation and Purification of B Cells from Human Peripheral Blood. J. Vis. Exp. (Pending Publication), e21523, doi: (2023).

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