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Generation of Murine Monoclonal Antibodies by Hybridoma Technology
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Immunology and Infection
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JoVE Journal Immunology and Infection
Generation of Murine Monoclonal Antibodies by Hybridoma Technology

Generation of Murine Monoclonal Antibodies by Hybridoma Technology

DOI:
10.3791/54832-v

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09:42 min

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January 02, 2017

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1Department of Biochemistry and Biology,University of Potsdam

Chapters

  • 00:05Title
  • 00:49Myeloma and Feeder Cell Culture Preparation
  • 02:41Cell Fusion
  • 05:58Hybridoma Cryoconservation, Polyclonal Limiting Dilution, and Mass Culture and Monoclonal Antibody Purification
  • 08:03Results: Representative Antibody Purification Analyses
  • 09:11Conclusion

Summary

Automatic Translation

Automatic Translation

An optimized protocol is presented for the generation of monoclonal antibodies based on the hybridoma technology. Mice were immunized with an immunoconjugate. Spleen cells were fused by PEG and an electric impulse with immortal myeloma cells. Antibody-producing hybridoma cells were selected by HAT and antigen-specific ELISA screening.

Tags

Keywords: Monoclonal AntibodiesHybridoma TechnologyMurineELISAFlow CytometryMyeloma CellsSplenocytesCell FusionFeeder Cells

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