AGL is supported by the NRF-CSUR (Grant Number CSUR60502163639) and the Centre for Tuberculosis Research from the South African Medical Research Council. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.ĭata Availability: All relevant data are within the manuscript and its Supporting Information files.įunding: This study was supported by Stellenbosch University Stellenbosch University Immunology Research Group and the Molecular Biology and Human Genetics Clinical Group National Research Foundation South African Medical Research Council. Received: SeptemAccepted: MaPublished: March 20, 2019Ĭopyright: © 2019 Moore et al. PLoS ONE 14(3):Įditor: Jeffrey Chalmers, The Ohio State University, UNITED STATES As such we suggest cell sorting as the preferred method for isolating pure B-cells to be used for downstream functional assays.Ĭitation: Moore DK, Motaung B, du Plessis N, Shabangu AN, Loxton AG, SU-IRG Consortium (2019) Isolation of B-cells using Miltenyi MACS bead isolation kits. These results illustrate the inefficiency of current available MACS B-cell isolation kits to produce pure B-cell populations, from which concrete findings can be made. Contamination of isolated B-cell fractions with platelets was the conclusive finding for all of the isolation techniques tested.
We used three Negative B-cell isolation kits as well as the Positive B-cell isolation kit from Miltenyi and compared the purity of each of the resulting B-cells fractions. 2) Positive selection–in which B-cells are labelled and actively removed from the sample. B-cells can be isolated by two main approaches: 1) Negative selection-in which B-cells remain “untouched” in their native state this is advantageous as it is likely that B-cells remain functionally unaltered by this process. Such populations are vital for studies investigating the functional capacity of B-cells, as the presence of other cell types may have indirect effects on B-cell function through cell-cell interactions or by secretion of several soluble molecules. This article describes the procedures used to isolate pure B-cell populations from whole blood using various Miltenyi magnetic-activated cell sorting (MACS) bead Isolation kits.
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