Question: What are the recommended T and B cell enrichment protocols?
Answer: Enrichment of T or B cells in a sample may be possible with the commercial products listed below. Note that most of these products have not been tested by 10x Genomics and so should be used as a starting point for optimization.
(i) Magnetic Beads
- EasySep Human T or B Cell Enrichment Kit* (STEMCELL Technologies)
- MACS Cell Separation with MicroBeads** (MACS Miltenyi Biotec)
- MagniSort™ Human T or B Cell Enrichment Kit* (Thermo Fisher Scientific)
- T Cell Enrichment Columns (R&D Systems)
(iii) Fluorescence-activated cell sorting (FACS)
* Protocols have not been tested by 10x Genomics for Single Cell V(D)J and are given as suggestions and may need to be optimized by the user.
** Magnetic MicroBeads are small (~50nm) and do not interfere during GEM generation. We have used CD3 MicroBeads to enrich for T cells in Clear Cell Renal Carcinoma and PBMC samples. After running through the V(D)J assay, T cell count enrichment improved by a factor of ~3.