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Isolation of Antigen-Specific, Antibody-Secreting Cells Using a Chip-Based Immunospot Array

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Human Monoclonal Antibodies

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1904))

Abstract

Antigen-specific monoclonal antibodies are useful tools to detect very small amounts of antigenic materials and are applicable for antibody therapeutics. To produce mouse monoclonal antibodies, a hybridoma between B lymphocytes and myeloma cells is used to produce antigen-specific monoclonal antibodies. However, a good hybridoma system is not available to obtain human monoclonal antibodies. To produce antigen-specific human monoclonal antibodies, transformation of B lymphocytes with Epstein-Barr viruses or a phage-display system is used. Here, we describe the screening of antigen-specific, antibody-secreting cells using microwell array chips to obtain antigen-specific human monoclonal antibodies. The system can be applied to screen antigen-specific, antibody-secreting cells from any animal species.

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References

  1. Kohler G, Milstein C (1975) Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256:495–497

    Article  CAS  Google Scholar 

  2. Kozbor D, Roder JC (1981) Requirements for the establishment of high-titered human monoclonal antibodies against tetanus toxoid using the Epstein-Barr virus technique. J Immunol 127:1275–1280

    CAS  PubMed  Google Scholar 

  3. Traggiai E, Becker S, Subbarao K, Kolesnikova L, Uematsu Y, Gismondo MR et al (2004) An efficient method to make human monoclonal antibodies from memory B cells: potent neutralization of SARS coronavirus. Nat Med 10:871–875

    Article  CAS  Google Scholar 

  4. McCafferty J, Griffiths AD, Winter G, Chiswell DJ (1990) Phage antibodies: filamentous phage displaying antibody variable domains. Nature 348:552–554

    Article  CAS  Google Scholar 

  5. Winter G, Griffiths AD, Hawkins RE, Hoogenboom HR (1994) Making antibodies by phage display technology. Annu Rev Immunol 12:433–455

    Article  CAS  Google Scholar 

  6. Ozawa T, Kinoshita K, Kadowaki S, Tajiri K, Kondo S, Honda R et al (2009) MAC-CCD system: a novel lymphocyte microwell-array chip system equipped with CCD scanner to generate human monoclonal antibodies against influenza virus. Lab Chip 9:158–163

    Article  CAS  Google Scholar 

  7. Tajiri K, Kishi H, Tokimitsu Y, Kondo S, Ozawa T, Kinoshita K et al (2007) Cell-microarray analysis of antigen-specific B-cells: single cell analysis of antigen receptor expression and specificity. Cytometry A 71:961–967

    Article  Google Scholar 

  8. Tokimitsu Y, Kishi H, Kondo S, Honda R, Tajiri K, Motoki K et al (2007) Single lymphocyte analysis with a microwell array chip. Cytometry A 71:1003–1010

    Article  Google Scholar 

  9. Yamamura S, Kishi H, Tokimitsu Y, Kondo S, Honda R, Rao SR et al (2005) Single-cell microarray for analyzing cellular response. Anal Chem 77:8050–8056

    Article  CAS  Google Scholar 

  10. Jin A, Ozawa T, Tajiri K, Obata T, Kondo S, Kinoshita K et al (2009) A rapid and efficient single-cell manipulation method for screening antigen-specific antibody-secreting cells from human peripheral blood. Nat Med 15:1088–1092

    Article  CAS  Google Scholar 

  11. Love JC, Ronan JL, Grotenbreg GM, van der Veen AG, Ploegh HL (2006) A microengraving method for rapid selection of single cells producing antigen-specific antibodies. Nat Biotechnol 24:703–707

    Article  CAS  Google Scholar 

  12. Deutsch M, Deutsch A, Shirihai O, Hurevich I, Afrimzon E, Shafran Y et al (2006) A novel miniature cell retainer for correlative high-content analysis of individual untethered non-adherent cells. Lab Chip 6:995–1000

    Article  CAS  Google Scholar 

  13. Biran I, Walt DR (2002) Optical imaging fiber-based single live cell arrays: a high-density cell assay platform. Anal Chem 74:3046–3054

    Article  CAS  Google Scholar 

  14. Zaimoku Y, Takamatsu H, Hosomichi K, Ozawa T, Nakagawa N, Imi T et al (2017) Identification of an HLA class I allele closely involved in the autoantigen presentation in acquired aplastic anemia. Blood 129:2908–2916

    Article  CAS  Google Scholar 

  15. Kurosawa N, Yoshioka M, Fujimoto R, Yamagishi F, Isobe M (2012) Rapid production of antigen-specific monoclonal antibodies from a variety of animals. BMC Biol 10:80

    Article  CAS  Google Scholar 

  16. Giudicelli V, Chaume D, Lefranc MP (2004) IMGT/V-QUEST, an integrated software program for immunoglobulin and T cell receptor V-J and V-D-J rearrangement analysis. Nucleic Acids Res 32:W435–W440

    Article  CAS  Google Scholar 

  17. Bernasconi NL, Traggiai E, Lanzavecchia A (2002) Maintenance of serological memory by polyclonal activation of human memory B cells. Science 298:2199–2202

    Article  CAS  Google Scholar 

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Acknowledgment

This work was supported by grants from the Toyama Medical Bio-Cluster Project and Hokuriku Innovation Cluster for Health Science of the Ministry of Education, Culture, Sports, Science and Technology, Japan. This work was also supported by JSPS KAKENHI Grant Number JP16H06499 (H.K.), the Platform Project for Supporting in Drug Discovery and Life Science Research (Platform for Drug Discovery, Informatics, and Structural Life Science) from the Ministry of Education, Culture, Sports, Science (MEXT) and the Japan Agency for Medical Research and Development (AMED) (A.M.), and the Platform Project for Supporting Drug Discovery and Life Science Research (Basis for Supporting Innovative Drug Discovery and Life Science Research (BINDS)) from AMED under Grant Number JP18am0101077 (T.O.).

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Correspondence to Hiroyuki Kishi .

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Kishi, H., Ozawa, T., Hamana, H., Kobayashi, E., Muraguchi, A. (2019). Isolation of Antigen-Specific, Antibody-Secreting Cells Using a Chip-Based Immunospot Array. In: Steinitz, M. (eds) Human Monoclonal Antibodies. Methods in Molecular Biology, vol 1904. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8958-4_6

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  • DOI: https://doi.org/10.1007/978-1-4939-8958-4_6

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8957-7

  • Online ISBN: 978-1-4939-8958-4

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