1. High-Throughput Mapping of B Cell Receptor Sequences to Antigen Specificity
- Author
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Charissa Oosthuysen, Andrea R. Shiakolas, Ivelin S. Georgiev, Priyamvada Acharya, Kelsey A. Pilewski, Rutendo E. Mapengo, Mark Connors, Daniel Lingwood, Ian Setliff, Lynn Morris, Juliana S. Qin, Barney S. Graham, Katarzyna Janowska, Amyn A. Murji, Allison R. Greenplate, Masaru Kanekiyo, Kevin J Kramer, Nagarajan Raju, Larance Ronsard, Simone I. Richardson, and Wyatt J. McDonnell
- Subjects
THP-1 Cells ,B-cell receptor ,Receptors, Antigen, B-Cell ,HIV Antibodies ,Biology ,Article ,General Biochemistry, Genetics and Molecular Biology ,Epitopes ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Antibody Repertoire ,Antigen ,Humans ,Antigens ,Cells, Cultured ,030304 developmental biology ,Systems immunology ,0303 health sciences ,breakpoint cluster region ,High-Throughput Nucleotide Sequencing ,Sequence Analysis, DNA ,Antibodies, Neutralizing ,Virology ,High-Throughput Screening Assays ,Vaccination ,HEK293 Cells ,biology.protein ,Single-Cell Analysis ,Antibody ,Epitope Mapping ,030217 neurology & neurosurgery - Abstract
B cell receptor (BCR) sequencing is a powerful tool for interrogating immune responses to infection and vaccination, but it provides limited information about the antigen specificity of the sequenced BCRs. Here, we present LIBRA-seq (linking B cell receptor to antigen specificity through sequencing), a technology for high-throughput mapping of paired heavy- and light-chain BCR sequences to their cognate antigen specificities. B cells are mixed with a panel of DNA-barcoded antigens so that both the antigen barcode(s) and BCR sequence are recovered via single-cell next-generation sequencing. Using LIBRA-seq, we mapped the antigen specificity of thousands of B cells from two HIV-infected subjects. The predicted specificities were confirmed for a number of HIV- and influenza-specific antibodies, including known and novel broadly neutralizing antibodies. LIBRA-seq will be an integral tool for antibody discovery and vaccine development efforts against a wide range of antigen targets.
- Published
- 2019