Cited by 294Monoclonal and Polyclonal Antibodies ResearchT-cell and B-cell Immunologyvaccines and immunoinformatics approaches

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Monoclonal antibodies isolated without screening by analyzing the variable-gene repertoire of plasma cells

Reddy, Sai T, Ge, Xin, Miklos, Aleksandr E, Hughes, Randall A, Kang, Seung Hyun, Hoi, Kam Hon, Chrysostomou, Constantine, Hunicke-Smith, Scott P, Iverson, Brent L, Tucker, Philip W, Ellington, Andrew D, Georgiou, George·2010·Nature Biotechnology

DOI 10.1038/nbt.1673

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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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Additional file 2 of Characterization of B cell receptor H-CDR3 repertoire of spleen in PRV-infected mice
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Additional file 1 of Characterization of B cell receptor H-CDR3 repertoire of spleen in PRV-infected mice
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Wu, H., Zhou, Z., Xie, S., Yan, R., Gong, M., Tian, X., & Wang, Z. (2022) · Figshare · doi:10.6084/m9.figshare.19311316
Additional file 9 of Similarity measurements of B cell receptor repertoire in baseline mice showed spectrum convergence of IgM
Wu, H., Zhou, Z., Xie, S., Yan, R., Gong, M., Tian, X., & Wang, Z. (2022) · Figshare · doi:10.6084/m9.figshare.19311340
Additional file 3 of Similarity measurements of B cell receptor repertoire in baseline mice showed spectrum convergence of IgM
Wu, H., Zhou, Z., Xie, S., Yan, R., Gong, M., Tian, X., & Wang, Z. (2022) · Figshare · doi:10.6084/m9.figshare.19311322
Additional file 6 of Similarity measurements of B cell receptor repertoire in baseline mice showed spectrum convergence of IgM
Wu, H., Zhou, Z., Xie, S., Yan, R., Gong, M., Tian, X., & Wang, Z. (2022) · Figshare · doi:10.6084/m9.figshare.19311331
Additional file 5 of Similarity measurements of B cell receptor repertoire in baseline mice showed spectrum convergence of IgM
Wu, H., Zhou, Z., Xie, S., Yan, R., Gong, M., Tian, X., & Wang, Z. (2022) · Figshare · doi:10.6084/m9.figshare.19311328
Additional file 4 of Similarity measurements of B cell receptor repertoire in baseline mice showed spectrum convergence of IgM
Wu, H., Zhou, Z., Xie, S., Yan, R., Gong, M., Tian, X., & Wang, Z. (2022) · Figshare · doi:10.6084/m9.figshare.19311325
Additional file 7 of Similarity measurements of B cell receptor repertoire in baseline mice showed spectrum convergence of IgM
Wu, H., Zhou, Z., Xie, S., Yan, R., Gong, M., Tian, X., & Wang, Z. (2022) · Figshare · doi:10.6084/m9.figshare.19311334
Genesis of immune diversity and selection of catalytic antibodies : a new investigation
Shahsavarian, M. A. (2015) · theses.fr (ABES) · doi:10.70675/da4996c0z7eafz499dz8c60zed99ba079c62
Convergent antibody signatures for the measles virus in transgenic rats expressing a human B cell IG repertoire
Dubois, A. (2015) · theses.fr (ABES) · doi:10.70675/b3c57c5cza980z4ea9zb000z900fae40cdfc
A la recherche d’anticorps humains et de nouveaux biomarqueurs pour le ciblage de la plaque d’athérome
Hémadou, A. (2017) · theses.fr (ABES) · doi:10.70675/1c2fc378zb806z46b2zafb9z47a32f1c994c
Sélection de fragments d’anticorps dirigés contre les microcystines pour la mise au point de tests d’immunodétection
Maalouf, R. (2018) · doi:10.70675/e40edc78z0f5ez4730z81e9z999ab53748b4
Statistical inference of evolutionary paths
Spisak, N. (2022) · theses.fr (ABES) · doi:10.70675/6f7a4d22z6762z4d3ezbc4czfd218ca968cc
Integrating computational and experimental approaches to identify antigen–specific antibodies
Abbate, M. F., Pereira, P., Toxe, P., Maestrali, N., Gagnaire, M., Vigne, E., Shahsavarian, M. A., Mora, T., & Walczak, A. M. (2026) · Frontiers in Immunology · doi:10.3389/fimmu.2026.1951538

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: Reddy, Sai T
Ge, Xin
Miklos, Aleksandr E
Hughes, Randall A
Kang, Seung Hyun
Hoi, Kam Hon
Iverson, Brent L
Ellington, Andrew D
Georgiou, George
Venue: Nature Biotechnology
Publisher: Springer Science and Business Media LLC
Published: 2010-08-29
Volume: 28
Issue: 9
Pages: 965-969
ISSN: 0733-222X (ISSN-L)
1087-0156
1546-1696
DOI: 10.1038/nbt.1673
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: Life Sciences › Immunology and Microbiology › Immunology › 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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Monoclonal antibodies isolated without screening by analyzing the variable-gene repertoire of plasma cells

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