2025-04-01
ENA first public
ERC000011
ENA-CHECKLIST
Animal Plant and Health Agency
INSDC center name
public
INSDC status
ERS22133941
SRA accession
E-MTAB-14606:HB099_GEX
Submitter Id
20
year
age
ArrayExpress
broker name
PBMCs
cell_type
not collected
collection date
human
common name
Protocols: PBMCs donated from three second dose donors at a single-cell level following 3-hour RBD-stimulation using the Cell Ranger (10x Genomics) were performed. Blood was obtained from three healthy control participants at least 3 weeks after their second COVID-19 vaccination, PBMCs were then obtained by Ficoll-Paque density centrifugation. PBMCs were plated in 96 well plates at 1 x 106 cells/mL in complete media (RPMI, 5% FCS, Pen/strep) supplemented with the following stimuli to 200 µL. For each participant there was one control well (+PBS), one RBD (500ng RBD - Biolegend), one peptide mix (500ng of Spike, Membrane and Nucleocapsid - Miltenyi PepTivator). Samples were incubated for 3 hours at 37 °C, at 2.5h 1 µL of hashtag antibodies (Total-Seq™-C, Biolegend) were added to each well to enable 10X genomics lane multiplexing on a per patient basis. Cells were collected and cooled to 4 °C and washed, 400g at 4 °C, in PBS+5% FCS and again in PBS+0.004% non-acetylated BSA. Samples were counted and pooled at equal numbers, washed again and resuspended in PBS+0.004% non-acetylated BSA to 1,000 cell/uL for single cell transcriptomics.  10X 5' single cell protocol 10X 5' single cell protocol
description
adult
developmental stage
None
disease
not collected
geographic location (country and/or sea)
2
individual
not applicable
isolate
Homo sapiens
organism
Homo sapiens
organism
blood
organism part
Homo sapiens
scientific_name
female
sex
42
day
time since 2nd dose
HB099_GEX
title