Monoclonal antibodies isolated without screening by analyzing the variable-gene repertoire of plasma cells
Abstract
Isolation of antigen-specific monoclonal antibodies (mAbs) and antibody fragments relies on high-throughput screening of immortalized B cells or recombinant antibody libraries. We bypassed the screening step by using high-throughput DNA sequencing and bioinformatic analysis to mine antibody variable region (V)-gene repertoires from bone marrow plasma cells (BMPC) of immunized mice. BMPCs, which cannot be immortalized, produce the vast majority of circulating antibodies. We found that the V-gene repertoire of BMPCs becomes highly polarized after immunization, with the most abundant sequences represented at frequencies between approximately 1% and >10% of the total repertoire. We paired the most abundant variable heavy (V(H)) and variable light (V(L)) genes based on their relative frequencies, reconstructed them using automated gene synthesis, and expressed recombinant antibodies in bacteria or mammalian cells. Antibodies generated in this manner from six mice, each immunized with one of three antigens were overwhelmingly antigen specific (21/27 or 78%). Those generated from a mouse with high serum titers had nanomolar binding affinities.
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A similar approach based on identifying highly abundant sequences was used to select antigen-specific molecules in immunized mice.
“A similar approach based on identifying highly abundant sequences was used to select antigen-specific molecules in immunized mice [216].”
From Computational approaches to therapeutic antibody design: established methods and emerging trends (2020) · cites this paper as [216]
Sequencing a sample of the repertoire and identifying newly abundant sequence-similar BCRs is used as a simple bioinformatic method to identify new antibodies specific for the antigen of interest.
“Sequencing a sample of the repertoire and identifying newly abundant sequence-similar BCRs is being used as a simple bioinformatic method to identify new antibodies specific for the antigen of interest.”
From Computational approaches to therapeutic antibody design: established methods and emerging trends (2020) · cites this paper as [216]
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- Authors
- Reddy, S. T., Ge, X., Miklos, A. E., Hughes, R. A., Kang, S. H., Hoi, K. H., Chrysostomou, C., Hunicke-Smith, S. P., Iverson, B. L., Tucker, P. W., Ellington, A. D., & Georgiou, G.
- Affiliations
- The University of Texas at Austin (US)
- Applied Research Laboratories, The University of Texas at Austin (US)
- Corresponding author
- Georgiou, George
- ORCID
- Venue
- Nature Biotechnology
- Published
- 2010-08-29
- Volume
- 28
- Issue
- 9
- Pages
- 965-969
- ISSN
- 0733-222X (ISSN-L)
- 1087-0156
- 1546-1696
- OpenAlex
- W1968091806
- Cited by
- 292
- References
- 32
- Citations by year
- 2026: 2 · 2025: 9 · 2024: 10 · 2023: 16 · 2022: 20 · 2021: 18 · 2020: 14 · 2019: 17 · 2018: 24 · 2017: 31 · 2016: 23 · 2015: 20 · 2014: 24 · 2013: 27 · 2012: 23
- FWCI
- 16.63
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Biotechnology (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Health Sciences › Medicine › Radiology, Nuclear Medicine and Imaging › Monoclonal and Polyclonal Antibodies Research
- Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › vaccines and immunoinformatics approaches
- Keywords
- monoclonal antibodies, antibody repertoire, plasma cells, antigen-specific antibodies, antibody fragments, immunization, antibody affinity, recombinant antibody production, next-generation sequencing
- MeSH terms
- Animals, Antibodies, Monoclonal, Antigens, Base Sequence, Humans, Immunization, Immunoglobulin Variable Region, Immunoglobulin Heavy Chains, Molecular Sequence Data, Plasma Cells, Complementarity Determining Regions, Mice, High-Throughput Screening Assays, High-Throughput Nucleotide Sequencing
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