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1.
Cell Rep ; 38(3): 110242, 2022 01 18.
Article in English | MEDLINE | ID: covidwho-1588137

ABSTRACT

Characterization of COVID-19 antibodies has largely focused on memory B cells; however, it is the antibody-secreting plasma cells that are directly responsible for the production of serum antibodies, which play a critical role in resolving SARS-CoV-2 infection. Little is known about the specificity of plasma cells, largely because plasma cells lack surface antibody expression, thereby complicating their screening. Here, we describe a technology pipeline that integrates single-cell antibody repertoire sequencing and mammalian display to interrogate the specificity of plasma cells from 16 convalescent patients. Single-cell sequencing allows us to profile antibody repertoire features and identify expanded clonal lineages. Mammalian display screening is used to reveal that 43 antibodies (of 132 candidates) derived from expanded plasma cell lineages are specific to SARS-CoV-2 antigens, including antibodies with high affinity to the SARS-CoV-2 receptor-binding domain (RBD) that exhibit potent neutralization and broad binding to the RBD of SARS-CoV-2 variants (of concern/interest).


Subject(s)
Antibodies, Neutralizing/isolation & purification , Plasma Cells/metabolism , SARS-CoV-2/immunology , Single-Cell Analysis/methods , Animals , Antibodies, Viral/isolation & purification , COVID-19/immunology , COVID-19/prevention & control , Cells, Cultured , Cohort Studies , Gene Library , HEK293 Cells , High-Throughput Nucleotide Sequencing/methods , Humans , Mammals , Neutralization Tests , Peptide Library , Plasma Cells/chemistry
2.
SoftwareX ; 15: 100710, 2021 Jul.
Article in English | MEDLINE | ID: covidwho-1281569

ABSTRACT

Lateral flow Point-Of-Care Tests (POCTs) are a valuable tool for rapidly detecting pathogens and the associated immune response in humans and animals. In the context of the SARS-CoV-2 pandemic, they offer rapid on-site diagnostics and can relieve centralized laboratory testing sites, thus freeing resources that can be focused on especially vulnerable groups. However, visual interpretation of the POCT test lines is subjective, error prone and only qualitative. Here we present pyPOCQuant, an open-source tool implemented in Python 3 that can robustly and reproducibly analyze POCTs from digital images and return an unbiased and quantitative measurement of the POCT test lines.

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