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Generation of Discriminative Human Monoclonal Antibodies from Rare Antigen-specific B Cells Circulating in Blood
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JoVE Journal Immunologia e infezioni
Generation of Discriminative Human Monoclonal Antibodies from Rare Antigen-specific B Cells Circulating in Blood

Generation of Discriminative Human Monoclonal Antibodies from Rare Antigen-specific B Cells Circulating in Blood

DOI:
10.3791/56508-v

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13:14 min

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February 06, 2018

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, , ,
1CRCINA, Inserm, CNRS, Université d’Angers,Université de Nantes, 2LabEx IGO, “Immunotherapy, Graft, Oncology”, 3CHU Nantes, France

Capitoli

  • 00:04Titolo
  • 01:17Isolation of Human Peripheral Blood Mononuclear Cells
  • 02:00Tetramer-Associated Magnetic Enrichment of Antigen Specific B-Cells
  • 03:58Staining of Antigen Specific Human B-Cells and Cell Sorting
  • 04:56Single Cell RT-PCR
  • 06:00Screening and Verification of Heavy and Light Chain Expressing Vectors
  • 07:45Production of Monoclonal Antibodies
  • 10:33Results: Detection of Relevant Antigen Specific B-Cells and Generation of Highly Discriminative Monoclonal Antibody from Donor Peripheral Blood Mononuclear Cells
  • 12:41Conclusion

Summary

Traduzione automatica

Traduzione automatica

We describe a method for the production of human antibodies specific for an antigen of interest, starting from rare B cells circulating in human blood. Generation of these natural antibodies is efficient and rapid, and the antibodies obtained can discriminate between highly related antigens.

Tags

Human Monoclonal AntibodiesAntigen-specific B CellsBloodDiscriminative AntibodiesImmunotherapyB LymphocytesDensity GradientMononuclear CellsTetramersMagnetic MicrobeadsCell Separation

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