Longitudinal scRNAseq profiling of memory B cells in mRNA vaccinated individuals post BA.1-breakthrough infection
Longitudinal scRNAseq profiling of memory B cells in mRNA vaccinated individuals post BA.1-breakthrough infection
For each sample, we always FACS sorted (MA900, Sony) an initial pool of 50.000 total peripheral CD3-CD14-CD56- CD19+ IgD- cells and afterward, to enrich for cells of interest, only sorted CD19+CD38low antibody secreting cells (ASCs), CD19hi IgD+ cells and SARS-CoV-2 Spike/RBD PE/tetramer positive B cells, leading to approximately 55000-60000 total sorted cells per sample. Inferred cell types per sample can be found in the corresponding sample_information_filtered.tsv file. Cells were counted and up to 20 000 cells were loaded in the 10x Chromium Controller to generate single-cell gel-beads in emulsion. After reverse transcription, gel-beads in emulsion were disrupted. Barcoded complementary DNA was isolated and amplified by PCR. The scRNA-seq libraries were generated using Chromium Next GEM Single Cell V(D)J Reagent Kit v.1.1 with Feature Barcoding (10x Genomics) according to the manufacturerâ€™s protocol. Gene expression (mRNA), ADT and VDJ BCR libraries were constructed.