GSM8239694: BWpool2-VDJ; Homo sapiens; OTHER
Single live CD45+ CD3+ cells were enriched from cryopreserved PBMC samples obtained pre-treatment and on-treatment at 4, 8, 12, 24, and 32 weeks by flow cytometry sorting using a BD FACSAria II (BD Biosciences). Cells were counted, hashed with TotalSeq-C0251, TotalSeq-C0252, TotalSeq-C0253 and TotalSeq-C0254 anti-human antibodies, BioLegend) and pooled in 2 separate batches across 3 GEMs (batch 1 = BWpool1 & BWpool2 containing PreTx/TotalSeq-C0251, OnTx.4wk/TotalSeq-C0252,  OnTx.8wk/TotalSeq-C0253,  OnTx.12wk/TotalSeq-C0254; batch2 = BWpool3 containing  OnTx.24wk/TotalSeq-C0251, OnTx.32wk/TotalSeq-C0252). scRNA/VDJ-seq (5'GEX, VDJ, HTO): The pooled cells were then counted again and immediately loaded onto the chromium chip G using the standard protocol for the Chromium single-cell 5' kit v2 (10x Genomics, Inc). Following the final bead purification, all three libraries were pooled as 80% mRNA library, 10% ADT library and 10% HTO library before sequencing.