The DOI matches its Crossref record; 32 of the 32 references it cites were found in a registry or an index.
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.
Overview
Integrity signals
Registry
Cites
Statistics
Verification
Claims the paper makes
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
In the citing paper
“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]
In the citing paper
“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]
Citations
Cites
Continuous cultures of fused cells secreting antibody of predefined specificity
Köhler, G., & Milstein, C. (1975) · Nature · doi:10.1038/256495a0 · Cited by 17,200
Next-generation DNA sequencing
Shendure, J., & Ji, H. (2008) · Nature Biotechnology · doi:10.1038/nbt1486 · Cited by 4,509
Brochet, X., Lefranc, M., & Giudicelli, V. (2008) · Nucleic Acids Research · doi:10.1093/nar/gkn316 · Cited by 1,220
Making antibody fragments using phage display libraries
Clackson, T., Hoogenboom, H. R., Griffiths, A. D., & Winter, G. P. (1991) · Nature · doi:10.1038/352624a0 · Cited by 1,211
Competence and competition: the challenge of becoming a long-lived plasma cell
Radbruch, A., Muehlinghaus, G., Luger, E., Inamine, A., Smith, K. G. C., Dörner, T., & Hiepe, F. (2006) · Nature reviews. Immunology · doi:10.1038/nri1886 · Cited by 1,033
Rapid cloning of high-affinity human monoclonal antibodies against influenza virus
Wrammert, J., Smith, K., Miller, J., Langley, W. A., Kokko, K. E., Larsen, C. P., Zheng, N., Mays, I., Garman, L., Helms, C., et al. (2008) · Nature · doi:10.1038/nature06890 · Cited by 1,026
Selecting and screening recombinant antibody libraries
Hoogenboom, H. R. (2005) · Nature Biotechnology · doi:10.1038/nbt1126 · Cited by 975
Regulation of plasma-cell development
Shelef, M. A., & Calame, K. L. (2005) · Nature reviews. Immunology · doi:10.1038/nri1572 · Cited by 917
Lifetime of plasma cells in the bone marrow
Manz, R. A., Thiel, A., & Radbruch, A. (1997) · Nature · doi:10.1038/40540 · Cited by 866
Traggiai, E., Becker, S., Subbarao, K., Kolesnikova, L., Uematsu, Y., Gismondo, M. R., Murphy, B. R., Rappuoli, R., & Lanzavecchia, A. (2004) · Nature Medicine · doi:10.1038/nm1080 · Cited by 697
MAINTENANCE OF SERUM ANTIBODY LEVELS
Manz, R. A., Hauser, A. E., Hiepe, F., & Radbruch, A. (2004) · Annual Review of Immunology · doi:10.1146/annurev.immunol.23.021704.115723 · Cited by 549
Feldhaus, M. J., Siegel, R. W., Opresko, L. K., Coleman, J. R., Feldhaus, J. M., Yeung, Y. A., Cochran, J. R., Heinzelman, P., Colby, D. W., Swers, J. S., et al. (2003) · Nature Biotechnology · doi:10.1038/nbt785 · Cited by 521
Krebber, A., Bornhauser, S., Burmester, J., Honegger, A., Willuda, J., Bosshard, H. R., & Plückthun, A. (1997) · Journal of Immunological Methods · doi:10.1016/s0022-1759(96)00208-6 · Cited by 492
A microengraving method for rapid selection of single cells producing antigen-specific antibodies
Love, J. C., Ronan, J. L., Grotenbreg, G. M., van der Veen, A. G., & Ploegh, H. L. (2006) · Nature Biotechnology · doi:10.1038/nbt1210 · Cited by 453
Glanville, J., Zhai, W., Berka, J., Telman, D., Huerta, G., Mehta, G. R., Ni, I., Mei, L., Sundar, P. D., Day, G. M. R., et al. (2009) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0909775106 · Cited by 440
High-Throughput Sequencing of the Zebrafish Antibody Repertoire
Weinstein, J. A., Jiang, N. J., White, R. A., III, Fisher, D. S., & Quake, S. R. (2009) · Science · doi:10.1126/science.1170020 · Cited by 432
High affinity germinal center B cells are actively selected into the plasma cell compartment
Phan, T. G., Paus, D., Chan, T. D., Turner, M. L., Nutt, S. L., Basten, A., & Brink, R. A. (2006) · The Journal of Experimental Medicine · doi:10.1084/jem.20061254 · Cited by 404
Boyd, S. D., Marshall, E. L., Merker, J. D., Maniar, J. M., Zhang, L. N., Sahaf, B., Jones, C. D., Simen, B. B., Hanczaruk, B., Nguyen, K. D., et al. (2009) · Science Translational Medicine · doi:10.1126/scitranslmed.3000540 · Cited by 387
Kwakkenbos, M. J., Diehl, S. A., Yasuda, E., Bakker, A. Q., van Geelen, C. M. M., Lukens, M. V., van Bleek, G. M., Widjojoatmodjo, M. N., Bogers, W. M. J. M., Mei, H. E., et al. (2009) · Nature Medicine · doi:10.1038/nm.2071 · Cited by 297
Jin, A., Ozawa, T., Tajiri, K., Obata, T., Kondo, S., Kinoshita, K., Kadowaki, S., Takahashi, K., Sugiyama, T., Kishi, H., et al. (2009) · Nature Medicine · doi:10.1038/nm.1966 · Cited by 261
Harvey, B. R., Georgiou, G., Hayhurst, A., Jeong, K. J., Iverson, B. L., & Rogers, G. K. (2004) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0400187101 · Cited by 217
Self-citationRibosome display: an in vitro method for selection and evolution of antibodies from libraries
Schaffitzel, C., Hanes, J., Jermutus, L., & Plückthun, A. (1999) · Journal of Immunological Methods · doi:10.1016/s0022-1759(99)00149-0 · Cited by 213
Isolation of engineered, full-length antibodies from libraries expressed in Escherichia coli
Mazor, Y., Blarcom, T. J. V., Mabry, R., Iverson, B. L., & Georgiou, G. (2007) · Nature Biotechnology · doi:10.1038/nbt1296 · Cited by 208
Self-citationThe changing economics of DNA synthesis
Carlson, R. H. (2009) · Nature Biotechnology · doi:10.1038/nbt1209-1091 · Cited by 166
Hayhurst, A., Happe, S., Mabry, R., Koch, Z., Iverson, B. L., & Georgiou, G. (2003) · Journal of Immunological Methods · doi:10.1016/s0022-1759(03)00100-5 · Cited by 146
Self-citationMeijer, P., Andersen, P. S., Hansen, M. H., Steinaa, L., Jensen, A. D., Lantto, J., Oleksiewicz, M. B., Tengbjerg, K., Poulsen, T. R., Coljee, V. W., et al. (2006) · Journal of Molecular Biology · doi:10.1016/j.jmb.2006.02.040 · Cited by 127
Gao, X. (2003) · Nucleic Acids Research · doi:10.1093/nar/gng143 · Cited by 118
A Focused Antibody Library for Improved Hapten Recognition
Persson, H., Lantto, J., & Ohlin, M. (2006) · Journal of Molecular Biology · doi:10.1016/j.jmb.2006.01.004 · Cited by 72
Antibody internalization studied using a novel IgG binding toxin fusion
Mazor, Y., Barnea, I., Keydar, I., & Benhar, I. (2007) · Journal of Immunological Methods · doi:10.1016/j.jim.2007.01.008 · Cited by 66
Synthetic Antibody Libraries Focused Towards Peptide Ligands
Cobaugh, C. W., Almagro, J. C., Pogson, M., Iverson, B. L., & Georgiou, G. (2008) · Journal of Molecular Biology · doi:10.1016/j.jmb.2008.02.037 · Cited by 63
Self-citationProtein fabrication automation
Cox, J. C., Lape, J., Sayed, M. A., & Hellinga, H. W. (2007) · Protein Science · doi:10.1110/ps.062591607 · Cited by 60
Rapid construction and characterization of synthetic antibody libraries without DNA amplification
Ge, X., Mazor, Y., Hunicke‐Smith, S., Ellington, A. D., & Georgiou, G. (2010) · Biotechnology and Bioengineering · doi:10.1002/bit.22712 · Cited by 33
Self-citation
Flagged first, then by citations
Cited by
Sorted by citations
Fusion protein linkers: Property, design and functionality
Chen, X., Zaro, J. L., & Shen, W. (2012) · Advanced Drug Delivery Reviews · doi:10.1016/j.addr.2012.09.039 · Cited by 2,157
The promise and challenge of high-throughput sequencing of the antibody repertoire
Georgiou, G., Ippolito, G. C., Beausang, J., Busse, C. E., Wardemann, H., & Quake, S. R. (2014) · Nature Biotechnology · doi:10.1038/nbt.2782 · Cited by 652
Self-citationB cells, plasma cells and antibody repertoires in the tumour microenvironment
Sharonov, G. V., Serebrovskaya, E. O., Yuzhakova, D. V., Britanova, O. V., & Chudakov, D. M. (2020) · Nature reviews. Immunology · doi:10.1038/s41577-019-0257-x · Cited by 630
Rogosch, T., Kerzel, S., Hoi, K. H., Zhang, Z., Maier, R. F., Ippolito, G. C., & Zemlin, M. (2012) · Frontiers in Immunology · doi:10.3389/fimmu.2012.00176 · Cited by 555
Self-citationBeyond natural antibodies: the power of in vitro display technologies
Bradbury, A. M., Sidhu, S. S., Dübel, S., & McCafferty, J. (2011) · Nature Biotechnology · doi:10.1038/nbt.1791 · Cited by 548
Metadata
- 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
- 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
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
