Skip to main content

Production of Human Monoclonal Antibodies by the Epstein–Barr Virus Method

  • Protocol
  • First Online:

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1060))

Abstract

Epstein–Barr virus (EBV) is a herpes virus which in vitro efficiently immortalizes nearly all human B lymphocytes. The lymphoblastoid diploid cell lines (LCL’s) thus generated preserve the characteristics of the cells initially infected by the virus: the cells produce and secrete immunoglobulins and also express these molecules on their surface. A selection of specific antibody-producing cells (i.e., antigen-committed cells) before EBV-infection or when LCL’s have already been established, enables isolation of monoclonal cell lines that secrete specific antibodies. If selection of antigen-committed cells is not feasible, secretion of specific antibodies by cloned LCL’s in limiting dilution cultures enables isolation of the desired cell lines. The method allows the production of human IgM, IgG, IgA, and IgE monoclonal antibodies from any individual. Monoclonal antibodies produced by the EBV method resemble the antibody repertoire of the donor of the lymphocytes. Human monoclonal antibodies are promising reagents for passive immunization.

This is a preview of subscription content, log in via an institution.

Buying options

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD   169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Nilsson K, Ponten J (1975) Classification and biological nature of established human hematopoietic cell lines. Int J Cancer 15:321–341

    Article  CAS  Google Scholar 

  2. Steinitz M, Klein G, Koskimies S, Mäkelä O (1977) EB virus-induced B lymphocyte cell lines producing specific antibody. Nature 269:420–422

    Article  CAS  Google Scholar 

  3. Kuppers R (2003) B cells under influence: transformation of B cells by Epstein-Barr virus. Nat Rev Immunol 3:801–812

    Article  Google Scholar 

  4. Nemerow GR, Wolfert R, McNaughton ME, Cooper NR (1985) Identification and characterization of the Epstein-Barr virus receptor on human lymphocytes-B and its relationship to the C3d complement receptor (Cr-2). J Virol 55:347–351

    Article  CAS  Google Scholar 

  5. Heath E, Begue-Pastor N, Chaganti S, Croom-Carter D, Shannon-Lowe C, Kube D, Feederle R, Delecluse HJ, Rickinson AB, Bell AI (2012) Epstein-Barr virus infection of naive B cells in vitro frequently selects clones with mutated immunoglobulin genotypes: implications for virus biology. PLoS Pathog 8(5):e1002697

    Article  CAS  Google Scholar 

  6. Ehlin-Henriksson B, Gordon J, Klein G (2003) B-lymphocyte subpopulations are equally susceptible to Epstein-Barr virus infection, irrespective of immunoglobulin isotype expression. Immunology 108:427–430

    Article  CAS  Google Scholar 

  7. Brown NA, Miller G (1982) Immunoglobulin expression by human lymphocytes-B clonally transformed by Epstein-Barr virus. J Immunol 128:24–29

    CAS  PubMed  Google Scholar 

  8. Miyawaki T, Kubagawa H, Butler JL, Cooper MD (1988) Ig isotypes produced by Ebv-transformed B-cells as a function of age and tissue distribution. J Immunol 140:3887–3892

    CAS  PubMed  Google Scholar 

  9. Stein LD, Chan MA, Hibi T, Dosch HM (1986) Epstein-Barr virus-induced Ige production in limiting dilution cultures of normal human B-cells. Eur J Immunol 16:1167–1170

    Article  CAS  Google Scholar 

  10. Steinitz M, Klein G (1980) Ebv-transformation of surface Iga-positive human-lymphocytes. J Immunol 125:194–196

    CAS  PubMed  Google Scholar 

  11. Shannon-Lowe C, Baldwin G, Feederle R, Bell A, Rickinson A, Delecluse HJ (2005) Epstein-Barr virus-induced B-cell transformation: quantitating events from virus binding to cell outgrowth. J Gen Virol 86:3009–3019

    Article  CAS  Google Scholar 

  12. Gil Y, Levy-Nabot S, Steinitz M, Laskov R (2007) Somatic mutations and activation-induced cytidine deaminase (AID) expression in established rheumatoid factor-producing lymphoblastoid cell line. Mol Immunol 44:494–505

    Article  CAS  Google Scholar 

  13. Chezar I, Lobel-Lavi L, Steinitz M, Laskov R (2008) Ongoing somatic hypermutation of the rearranged VH but not of the V-lambda gene in EBV-transformed rheumatoid factor-producing lymphoblastoid cell line. Mol Immunol 46:80–90

    Article  CAS  Google Scholar 

  14. Laskov R, Yahud V, Hamo R, Steinitz M (2011) Preferential targeting of somatic hypermutation to hotspot motifs and hypermutable sites and generation of mutational clusters in the IgVH alleles of a rheumatoid factor producing lymphoblastoid cell line. Mol Immunol 48:733–745

    Article  CAS  Google Scholar 

  15. Miller G, Lipman M (1973) Comparison of yield of infectious virus from clones of human and simian lymphoblastoid lines transformed by Epstein-Barr-virus. J Exp Med 138:1398–1412

    Article  CAS  Google Scholar 

  16. Steinitz M, Tamir S, Frodin JE, Lefvert AK, Mellstedt H (1988) Human monoclonal anti-idiotypic antibodies.1. Establishment of immortalized cell-lines from a tumor patient treated with mouse monoclonal-antibodies. J Immunol 141:3516–3522

    CAS  PubMed  Google Scholar 

  17. Traggiai E, Becker S, Subbarao K, Kolesnikova L, Uematsu Y, Gismondo MR, Murphy BR, Rappuoli R, Lanzavecchia A (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 

Download references

Acknowledgment

The help of Dr. A. Mahler in preparing the manuscript is greatly appreciated.

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2014 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Steinitz, M. (2014). Production of Human Monoclonal Antibodies by the Epstein–Barr Virus Method. In: Steinitz, M. (eds) Human Monoclonal Antibodies. Methods in Molecular Biology, vol 1060. Humana, Totowa, NJ. https://doi.org/10.1007/978-1-62703-586-6_6

Download citation

  • DOI: https://doi.org/10.1007/978-1-62703-586-6_6

  • Published:

  • Publisher Name: Humana, Totowa, NJ

  • Print ISBN: 978-1-62703-585-9

  • Online ISBN: 978-1-62703-586-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics