Skip to main content
SpringerLink
Log in

An Overview of Mouse Monoclonal Antibody Production

  • Protocol
  • First Online:
Protocols for the Diagnosis of Pig Viral Diseases

Part of the book series: Springer Protocols Handbooks ((SPH))

  • 1145 Accesses

Abstract

Antibodies are an important tool in the field of diagnostics and therapeutics owing to their affinity to bind with specific antigen. These can be of two types: polyclonal antibodies that are produced by a mixture of various B lymphocyte clones, and monoclonal antibodies (mAbs) which are secreted by a single clone of B lymphocytes. mAbs have a higher affinity to the target protein and are highly selective in nature which makes them the best choice for specific purposes. Since the development of hybridoma technology in 1975, a number of mAbs have been developed and are in use for therapeutic and diagnostic purposes. Besides the recent advances in high throughput mAb generation technologies, hybridoma is the most preferred method due to its nature to maintain the inherent antibody structure and functional information. This chapter focuses on the basics of hybridoma technology including various steps involved in production of mAbs and various critical points which must be considered while generation of monoclonal antibodies.

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

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 249.99
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

Institutional subscriptions

References

  1. Avrameas S (1983) Enzyme immunoassays and related techniques: development and limitations. In: Bachmann PA (ed) New developments in diagnostic virology. Springer-Verlag, New York, NY, pp 93–99

    Chapter  Google Scholar 

  2. Yolken RH (1983) Use of monoclonal antibodies for viral diagnosis. Curr Top Microbial Immunol 104:177–l95

    CAS  Google Scholar 

  3. Köhler G, Milstein C (1975) Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256:495–497. https://doi.org/10.1038/256495a0

    Article  PubMed  Google Scholar 

  4. Pandey S (2010) Hybridoma technology for production of monoclonal antibodies. Int J Pharm Sci Rev Res 1(2):88–94

    CAS  Google Scholar 

  5. Alberts B, Johnson A, Lewis J et al (2002) B cells and antibodies. In: Molecular biology of the cell, 4th edn. Garland Science, New York

    Google Scholar 

  6. Leenaars M, Hendriksen CF (2005) Critical steps in the production of polyclonal and monoclonal antibodies: evaluation and recommendations. ILAR J 46(3):269–279

    Article  CAS  Google Scholar 

  7. Potgieter LN, Ajidagba PA (1989a) Quantitation of canine distemper virus and antibodies by enzyme-linked immunosorbent assays using protein a and monoclonal antibody capture. J Vet Diagn Investig 1:110–115

    Article  CAS  Google Scholar 

  8. Singh RP, Bandyopadhyay SK, Sreenivasa BP, Dhar P (2004) Production and characterization of monoclonal antibodies to pests des Petitis Ruminanats (PPR) virus. Vet Res Commun 28(7):623–639

    Article  CAS  Google Scholar 

  9. Liu Y, Hao L, Li X, Wang L, Zhang J, Deng J, Tian K (2017) Development and characterization of canine distemper virus monoclonal antibodies. Monoclon Antib Immunodiagn Immunother 36:119–123

    Article  CAS  Google Scholar 

  10. Masuda M, Sato H, Kamata H, Katsuo T, Takenaka A, Miura R, Yoneda M, Tsukiyama-Kohara K, Mizumoto K, Kai C (2006) Characterization of monoclonal antibodies directed against the canine distemper virus nucleocapsid protein. Comp Immunol Microbiol Infect Dis 29:157–165

    Article  Google Scholar 

  11. Parray HA, Shukla S, Samal S, Shrivastava T, Ahmed S, Sharma C, Kumar R (2020) Hybridoma technology a versatile method for isolation of monoclonal antibodies, its applicability across species, limitations, advancement and future perspectives. Int Immunopharmacol 85:106639

    Article  CAS  Google Scholar 

  12. Lerner EA (1981) How to make a hybridoma. Yale J Biol Med 54(5):387–402

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Hundekar YR, Chimkode RM, Salagare MV (2017) Hybridoma technology: a new scientific tool for AIDS recovery. Int J Pharm Pharm Sci 9(4):1–16

    Article  CAS  Google Scholar 

  14. Greenfield EA (2018) Screening for good batches of fetal bovine serum for myeloma and Hybridoma growth. Cold Spring Harb Protoc 1(3). https://doi.org/10.1101/pdb.prot103150

  15. Fuller SA, Takahashi M, Hurrell JGR (2001) Cloning of Hybridoma cell lines by limiting dilution. Curr Protoc Mol Biol; Chapter 11:Unit11.8. https://doi.org/10.1002/0471142727.mb1108s01

  16. Halder H, Embelton M, Fischer R, de Geus B, Hendriksen C, de Leeuw W, Marx U, Balls M (1998) Comments on appendix C of the National Institutes of Health response to the petition of the American anti-vivisection society to prohibit the use of animals in the production of monoclonal antibodies. Altern Lab Anim 26(4):549–554

    Article  CAS  Google Scholar 

  17. Seppala IJT, Sarvas H, Peterfy H, Makala O (1981) The four subclasses of IgG can be isolated from mouse serum by using protein A-Sepharose. Scand J Immunol 14:335–342

    Article  CAS  Google Scholar 

  18. Cole SR, Ashman LK, Ey PL (1987) Biotinylation: an alternative to radioiodination for the identification of cell surface antigens in immunoprecipitates. Mol Immunol 24:699

    Article  CAS  Google Scholar 

  19. Schuh R, Kremmer E, Ego E, Wasiliu M, Thierfeldcr S (1992) Determination of monoclonal antibody specificity by immunoadsorption and Western blotting. J Immunol Methods 152:9–67

    Article  Google Scholar 

  20. Bi Z, Xia X, Wang Y, Mei Y (2015) Development and characterization of neutralizing monoclonal antibodies against canine distemper virus hemagglutinin protein. Microbiol Immunol 59:202–208

    Article  CAS  Google Scholar 

  21. An DJ, Kim TY, Song DS, Kang BK, Park BK (2008) An immunochromatography assay for rapid antemortem diagnosis of dogs suspected to have canine distemper. J Virol Methods 147:244–249

    Article  CAS  Google Scholar 

  22. Li Y, Cheng S, Xu H, Wang J, Cheng Y, Yang S, Luo B (2012) Development of a combined canine distemper virus specific RT-PCR protocol for the differentiation of infected and vaccinated animals (DIVA) and genetic characterization of the hemagglutinin gene of seven Chinese strains demonstrated in dogs. J Virol Methods 179:281–287

    Article  Google Scholar 

  23. Zhang Y, Xu G, Zhang L, Zhao J, Ji P, Li Y, Liu B, Zhang J, Zhao Q, Sun Y, Zhou E (2020) Development of a double monoclonal antibody–based sandwich enzyme-linked immunosorbent assay for detecting canine distemper virus. Appl Microbiol Biotechnol 104:10725–10735

    Article  CAS  Google Scholar 

  24. Ogino T, Wang X, Ferrone S (2003) Modified flow cytometry and cell-ELISA methodology to detect HLA class I antigen processing machinery components in cytoplasm and endoplasmic reticulum. J Immunol Methods 278:33–44

    Article  CAS  Google Scholar 

  25. Giraudon P, Wild TF (1981) Differentiation of measles virus strains and a strain of canine distemper virus by monoclonal antibodies. J Gen Virol 57:179–183

    Article  CAS  Google Scholar 

  26. Russell PH, Alexander DJ (1983) Antigenic variation of Newcastle disease virus strains detected by monoclonal antibodies. Arch Virol 75(4):243–253

    Article  CAS  Google Scholar 

  27. Sheshberadaran H, Chen SN, Norrby E (1983) Monoclonal antibodies against five structural components of measles virus. I. Characterisation of antigenic determinants on nine strains of measles virus. Virology 128:341–353

    Article  CAS  Google Scholar 

  28. Ter Meulen V, Loffler S, Carter MJ, Stephenson JR (1981) Antigenic characterization of measles and SSPE virus Haemagglutinin by monoclonal antibodies. J Gen Virol 57:357–364

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Division of Biological Products, ICAR-IVRI, Izatnagar, India

    Kaushal Kishor Rajak,  Kiran, Arfa Fayaz, Vishal Rai, Monu Karki, Chris Einstein, Ashok Kumar & R. P. Singh

  2. ICAR-National Organic Farming Research Institute, Tadong, Sikkim, India

    Mukesh Bhatt

  3. ICAR-National Research Centre on Pig, Rani, Guwahati, Assam, India

    Ajay Kumar Yadav

Authors
  1. Kaushal Kishor Rajak
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kiran
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Arfa Fayaz
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Vishal Rai
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Monu Karki
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Chris Einstein
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Mukesh Bhatt
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Ashok Kumar
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Ajay Kumar Yadav
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. R. P. Singh
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. National Research Centre on Pig, Indian Council of Agricultural Research, Guwahati, Assam, India

    Rajib Deb

  2. National Research Centre on Pig, Indian Council of Agricultural Research, Guwahati, Assam, India

    Ajay Kumar Yadav

  3. National Research Centre on Pig, Indian Council of Agricultural Research, Guwahati, Assam, India

    Swaraj Rajkhowa

  4. College of Animal Biotechnology, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab, India

    Yashpal Singh Malik

Rights and permissions

Reprints and permissions

Copyright information

© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Rajak, K.K. et al. (2022). An Overview of Mouse Monoclonal Antibody Production. In: Deb, R., Yadav, A.K., Rajkhowa, S., Malik, Y.S. (eds) Protocols for the Diagnosis of Pig Viral Diseases. Springer Protocols Handbooks. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2043-4_19

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-2043-4_19

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2042-7

  • Online ISBN: 978-1-0716-2043-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation