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Generation of Discriminative Human Monoclonal Antibodies from Rare Antigen-specific B Cells Circulating in Blood
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Chapters
- 00:04Title
- 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
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.
