Rapid Generation of Monoclonal Antibodies from Single B Cells by Ecobody Technology

Cell-free protein synthesis: Applications come of age

Carlson, E. D., Gan, R., Hodgman, C. E., & Jewett, M. C. (2011) · Biotechnology Advances · doi:10.1016/j.biotechadv.2011.09.016 · Cited by 707 · see citing works

Cell‐Free Protein Synthesis: Pros and Cons of Prokaryotic and Eukaryotic Systems

Zemella, A., Thoring, L., Hoffmeister, C., & Kubick, S. (2015) · ChemBioChem · doi:10.1002/cbic.201500340 · Cited by 260 · see citing works

Highly efficient recovery of functional single-chain Fv fragments from inclusion bodies overexpressed in Escherichia coli by controlled introduction of oxidizing reagent—application to a human single-chain Fv fragment

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A simplified and robust protocol for immunoglobulin expression in E scherichia coli cell‐free protein synthesis systems

Qi, C., Hanson, J., Steiner, A. R., Tran, C., Masikat, M. R., Chen, R., Zawada, J. F., Sato, A. K., Hallam, T. J., & Yin, G. (2015) · Biotechnology Progress · doi:10.1002/btpr.2082 · Cited by 105 · see citing works

Single B cell antibody technologies

Tiller, T. (2011) · New Biotechnology · doi:10.1016/j.nbt.2011.03.014 · Cited by 88 · see citing works

Generation of Recombinant Monoclonal Antibodies from Immunised Mice and Rabbits via Flow Cytometry and Sorting of Antigen-Specific IgG+ Memory B Cells

Starkie, D., Compson, J. E., Rapecki, S., & Lightwood, D. J. (2016) · PLoS ONE · doi:10.1371/journal.pone.0152282 · Cited by 69 · see citing works

Engineering toward a bacterial “endoplasmic reticulum” for the rapid expression of immunoglobulin proteins

Groff, D., Armstrong, S. M., Rivers, P. J., Zhang, J., Yang, J., Green, E., Rozzelle, J., Liang, S.-W., Kittle, J. D., Steiner, A. R., et al. (2014) · mAbs · doi:10.4161/mabs.28172 · Cited by 66 · see citing works

A Highly Controllable Reconstituted Cell-Free System -a Breakthrough in Protein Synthesis Research

Ohashi, H., Kanamori, T., Shimizu, Y., & Ueda, T. (2010) · Current Pharmaceutical Biotechnology · doi:10.2174/138920110791111889 · Cited by 65 · see citing works

N-terminal SKIK peptide tag markedly improves expression of difficult-to-express proteins in Escherichia coli and Saccharomyces cerevisiae

Ojima-Kato, T., Nagai, S., & Nakano, H. (2017) · Journal of Bioscience and Bioengineering · doi:10.1016/j.jbiosc.2016.12.004 · Cited by 65 · see citing works

A Robust High Throughput Platform to Generate Functional Recombinant Monoclonal Antibodies Using Rabbit B Cells from Peripheral Blood

Seeber, S., Ros, F., Thorey, I. S., Tiefenthaler, G., Kaluza, K., Lifke, V., Fischer, J. A. A., Klostermann, S., Endl, J., Kopetzki, E., et al. (2014) · PLoS ONE · doi:10.1371/journal.pone.0086184 · Cited by 62 · see citing works

Rapid production of antigen-specific monoclonal antibodies from a variety of animals

Kurosawa, N., Yoshioka, M., Fujimoto, R., Yamagishi, F., & Isobe, M. (2012) · BMC Biology · doi:10.1186/1741-7007-10-80 · Cited by 59 · see citing works

Rapid isolation of dengue-neutralizing antibodies from single cell-sorted human antigen-specific memory B-cell cultures

Cox, K. S., Tang, A., Chen, Z., Horton, M. S., Yan, H., Wang, X.-M., Dubey, S. A., DiStefano, D. J., Ettenger, A., Fong, R. H., et al. (2015) · mAbs · doi:10.1080/19420862.2015.1109757 · Cited by 48 · see citing works

Ecobody technology: Rapid monoclonal antibody screening method from single B cells using cell-free protein synthesis for antigen-binding fragment formation

Ojima-Kato, T., Nagai, S., & Nakano, H. (2017) · Scientific Reports · doi:10.1038/s41598-017-14277-0 · Cited by 44 · see citing works

Development and characterization of monoclonal antibodies against Shiga toxin 2 and their application for toxin detection in milk

He, X., McMahon, S., Skinner, C. B., Merrill, P., Scotcher, M. C., & Stanker, L. H. (2012) · Journal of Immunological Methods · doi:10.1016/j.jim.2012.12.005 · Cited by 40 · see citing works

‘Zipbody’ leucine zipper-fused Fab in<i>E. coli in vitro</i>and<i>in vivo</i>expression systems

Ojima-Kato, T., Fukui, K., Yamamoto, H., Hashimura, D., Miyake, S., Hirakawa, Y., Yamasaki, T., Kojima, T., & Nakano, H. (2016) · Protein Engineering Design and Selection · doi:10.1093/protein/gzw001 · Cited by 35 · see citing works

DNA repair capacity measured by high throughput alkaline comet assays in EBV-transformed cell lines and peripheral blood cells from cancer patients and healthy volunteers

Iwakawa, M., Goto, M., Noda, S., Sagara, M., Yamada, S., Yamamoto, N., Kawakami, Y., Matsui, Y., Miyazawa, Y., Yamazaki, H., et al. (2005) · Mutation Research/Genetic Toxicology and Environmental Mutagenesis · doi:10.1016/j.mrgentox.2005.07.012 · Cited by 35 · see citing works

In vitro generation of rabbit anti-Listeria monocytogenes monoclonal antibody using single cell based RT-PCR linked cell-free expression systems

Ojima-Kato, T., Hashimura, D., Kojima, T., Minabe, S., & Nakano, H. (2015) · Journal of Immunological Methods · doi:10.1016/j.jim.2015.10.001 · Cited by 31 · see citing works

Crystal structure of a conformation-dependent rabbit IgG Fab specific for amyloid prefibrillar oligomers

Arai, H., Glabe, C. C., & Luecke, H. (2012) · Biochimica et Biophysica Acta (BBA) - General Subjects · doi:10.1016/j.bbagen.2012.08.016 · Cited by 26 · see citing works

A novel strategy for generation of monoclonal antibodies from single B cells using rt-PCR technique and in vitro expression

Jiang, X. P., Suzuki, H., Hanai, Y., Wada, F., Hitomi, K., Yamane, T., & Nakano, H. (2006) · Biotechnology Progress · doi:10.1021/bp060092h · Cited by 25 · see citing works

In vitro generation of anti-hepatitis B monoclonal antibodies from a single plasma cell using single-cell RT-PCR and cell-free protein synthesis

Sabrina, Y., Ali, M., & Nakano, H. (2010) · Journal of Bioscience and Bioengineering · doi:10.1016/j.jbiosc.2009.07.001 · Cited by 10 · see citing works

Evolution of Escherichia coli Expression System in Producing Antibody Recombinant Fragments

Sandomenico, A., Sivaccumar, J. P., & Ruvo, M. (2020) · International Journal of Molecular Sciences · doi:10.3390/ijms21176324 · Cited by 134 · see citing works

Cell-Free Gene Expression: Methods and Applications

Hunt, A. C., Rasor, B. J., Seki, K., Ekas, H. M., Warfel, K. F., Karim, A. S., & Jewett, M. C. (2024) · Chemical Reviews · doi:10.1021/acs.chemrev.4c00116 · Cited by 116 · see citing works

A rapid cell-free expression and screening platform for antibody discovery

Hunt, A. C., Vögeli, B., Hassan, A. O., Guerrero, L., Kightlinger, W., Yoesep, D. J., Krüger, A., DeWinter, M. A., Diamond, M., Karim, A. S., et al. (2023) · Nature Communications · doi:10.1038/s41467-023-38965-w · Cited by 72 · see citing works

Monoclonal Antibodies for Bacterial Pathogens: Mechanisms of Action and Engineering Approaches for Enhanced Effector Functions

Vacca, F., Sala, C., & Rappuoli, R. (2022) · Biomedicines · doi:10.3390/biomedicines10092126 · Cited by 58 · see citing works

Full-length recombinant antibodies from Escherichia coli : production, characterization, effector function (Fc) engineering, and clinical evaluation

Rashid, M. H. (2022) · mAbs · doi:10.1080/19420862.2022.2111748 · Cited by 47 · see citing works

Advances in recombinant protein production in microorganisms and functional peptide tags

Ojima‐Kato, T. (2024) · Bioscience Biotechnology and Biochemistry · doi:10.1093/bbb/zbae147 · Cited by 25 · see citing works

Discovery and characterization of high-affinity, potent SARS-CoV-2 neutralizing antibodies via single B cell screening

Schardt, J. S., Pornnoppadol, G., Desai, A. A., Park, K. S., Zupancic, J. M., Makowski, E. K., Smith, M. D., Chen, H., Barbosa, M. G. D. M., Cascalho, M., et al. (2021) · Scientific Reports · doi:10.1038/s41598-021-99401-x · Cited by 23 · see citing works

Point-of-Care Peptide Hormone Production Enabled by Cell-Free Protein Synthesis

DeWinter, M. A., Thames, A. H., Guerrero, L., Kightlinger, W., Karim, A. S., & Jewett, M. C. (2023) · ACS Synthetic Biology · doi:10.1021/acssynbio.2c00680 · Cited by 20 · see citing works

A platform-agnostic, function first-based antibody discovery strategy using plasmid-free mammalian expression of antibodies

Zhang, R., Prabakaran, P., Yu, X., Mackness, B. C., Boudanova, E., Hopke, J., Sancho, J., Saleh, J., Cho, H., Zhang, N., et al. (2021) · mAbs · doi:10.1080/19420862.2021.1904546 · Cited by 13 · see citing works

A high-throughput, automated, cell-free expression and screening platform for antibody discoveryPreprint

Hunt, A. C., Vögeli, B., Kightlinger, W., Yoesep, D. J., Krüger, A., & Jewett, M. C. (2021) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2021.11.04.467378 · Cited by 12 · see citing works

Development of Porcine Monoclonal Antibodies with In Vitro Neutralizing Activity against Classical Swine Fever Virus from C-Strain E2-Specific Single B Cells

Wang, L., Madera, R., Li, Y., Gladue, D. P., Borca, M. V., McIntosh, M. T., & Shi, J. N. (2023) · Viruses · doi:10.3390/v15040863 · Cited by 11 · see citing works

Monoclonal Antibodies: A Prospective and Retrospective View

Sivaccumar, J., Sandomenico, A., Vitagliano, L., & Ruvo, M. (2021) · Current Medicinal Chemistry · doi:10.2174/0929867327666200219142231 · Cited by 14 · see citing works

Live long and active: Polypeptide-mediated assembly of antibody variable fragments

Lee, C., Choi, M., & MacKay, J. A. (2020) · Advanced Drug Delivery Reviews · doi:10.1016/j.addr.2020.10.017 · Cited by 9 · see citing works

Development of a dual monoclonal antibody sandwich enzyme-linked immunosorbent assay for the detection of swine influenza virus using rabbit monoclonal antibody by Ecobody technology

Sila-on, D., Chertchinnapa, P., Shinkai, Y., Kojima, T., & Nakano, H. (2020) · Journal of Bioscience and Bioengineering · doi:10.1016/j.jbiosc.2020.03.003 · Cited by 7 · see citing works

Toward the commercialization of recombinant pharmaceuticals expressed in plants

Washida, H., Yoshinaka, K., Yamada, O., Ookawa, S., & Yuki, M. (2025) · Plant Biotechnology · doi:10.5511/plantbiotechnology.24.1130a · Cited by 4 · see citing works

A Microfluidic Strategy to Capture Antigen‐Specific High‐Affinity B Cells

Alhassan, A. M., Shirure, V. S., Luo, J. J., Nguyen, B. B., Rollins, Z. A., Shergill, B. S., Zhu, X. D., Baumgarth, N., & George, S. C. (2024) · Advanced NanoBiomed Research · doi:10.1002/anbr.202300101 · Cited by 3 · see citing works

Expression of soluble recombinant human matrix metalloproteinase 9 and generation of its monoclonal antibody

Jeong, H., Kim, E., Kim, J.-K., Kim, Y., Lee, C., Ko, B. J., & Kim, B. (2021) · Protein Expression and Purification · doi:10.1016/j.pep.2021.105931 · Cited by 2 · see citing works

Investigation of Intestinal Bacteria-binding Monoclonal Antibodies Derived from Rabbit Single B CellsPreprint

Anwar, K., Kaneko, S., Kawakita, I., Adinegoro, D. S., Kihara, M., Ojima‐Kato, T., & Nakano, H. (2024) · bioRxiv (Cold Spring Harbor Laboratory) · doi:10.1101/2024.07.26.605397

Analysis and Application of Translation-Enhancing Peptides for Improved Production of Proteins Containing Polyproline

Yoshino, A., Shimoji, R., Nishikawa, Y., Nakano, H., & Ojima‐Kato, T. (2025) · SynBio · doi:10.3390/synbio3040014

Development of monoclonal antibody targeting membrane protein utilizing modified Ecobody technology

Adinegoro, D. S., Ojima‐Kato, T., & Nakano, H. (2026) · Journal of Bioscience and Bioengineering · doi:10.1016/j.jbiosc.2025.12.001

A rapid cell-free expression platform for screening of nanobody mutants

Zhang, H., Chen, M., Li, Y., He, Q., & Tu, Z. (2026) · Biotechnology and Bioprocess Engineering · doi:10.1007/s12257-026-00273-7

Identification of antibodies by phage display for immunohistochemical applications

Mairaville, C. (2021) · doi:10.70675/bbe469baz5631z452fzac83z2f6eaab87007

Identification and characterization of a high-affinity antigen-binding fragment against intestinal bacteria from non-immunized rabbit single B cells using Ecobody technology

Anwar, K., Kaneko, S., Kawakita, I., Adinegoro, D. S., Kihara, M., Kojima, T., Ojima‐Kato, T., & Nakano, H. (2026) · Journal of Bioscience and Bioengineering · doi:10.1016/j.jbiosc.2026.09.001

Ecobody technology: Rapid monoclonal antibody screening method from single B cells using cell-free protein synthesis for antigen-binding fragment formation

Ojima-Kato, T., Nagai, S., & Nakano, H. (2017) · Scientific Reports · doi:10.1038/s41598-017-14277-0 · Cited by 44 · see citing works

Ecobody technology: a novel antibody screening and production method of monoclonal antibodies by single B-cell RT-PCR and Escherichia coli in vivo and in vitro expression systems

Nakano, H., Kato, T., Morishita, S., Uchida, Y., Ritthisan, P., & Kojima, T. (2017) · The Japanese Biochemical Society/The Molecular Biology Society of Japan

A novel strategy for generation of monoclonal antibodies from single B cells using rt-PCR technique and in vitro expression

Jiang, X. P., Suzuki, H., Hanai, Y., Wada, F., Hitomi, K., Yamane, T., & Nakano, H. (2006) · Biotechnology Progress · doi:10.1021/bp060092h · Cited by 25 · see citing works

Development of a CHO-Based Cell-Free Platform for Synthesis of Active Monoclonal Antibodies

Martin, R. W., Majewska, N. I., Chen, C. X., Albanetti, T., Jimenez, R. B., Schmelzer, A. E., Jewett, M. C., & Roy, V. (2017) · ACS Synthetic Biology · doi:10.1021/acssynbio.7b00001 · Cited by 104 · see citing works

Studies on a rapid monoclonal antibody generation using single-cell RT-PCR and cell-free protein synthesis

Yunita, S. (2011)