Rapid purification of billions of circulating CD19+ B cells directly from leukophoresis samples

N Biotechnol. 2018 Nov 25:46:14-21. doi: 10.1016/j.nbt.2018.05.006. Epub 2018 Jun 2.

Abstract

The study of the biology and function of B cells, or the dissection and in vitro creation of enormous recombinant antibody repertoires, requires the isolation of large numbers of pure CD19+ B cells. The StraightFrom® Leukopak CD19 MicroBead Kit was recently introduced as a fast and robust kit to isolate human CD19+ B cells. This uses paramagnetic microbeads conjugated to high-affinity anti-CD19 monoclonal antibodies to bind B cells in leukapheresis (Leukopak) samples. The overall purity of the isolated cells, together with the characterization of the different CD19+ subclasses, was assessed by flow cytometry using a recombinant (REAffinity) antibody panel, revealing that the method allowed the recovery of over 93% of CD19+ B cells without any pre-purification step. This enables the relatively straightforward purification of all the circulating CD19+ B cells in a single donor.

Keywords: B cells; Magnetic bead purification; Recombinant antibodies.

MeSH terms

  • Antibodies, Monoclonal / immunology
  • Antigens, CD19 / immunology
  • Antigens, CD19 / metabolism*
  • B-Lymphocytes / cytology*
  • B-Lymphocytes / immunology
  • B-Lymphocytes / metabolism*
  • Cell Separation*
  • Flow Cytometry*
  • Humans

Substances

  • Antibodies, Monoclonal
  • Antigens, CD19