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Chapters
- 00:05Title
- 01:41Staining and Sorting PBMCs for CD22+ and IgG+ Cells
- 02:46Plating the Sorted PBMCs Including EBV Infection
- 04:13An ELISA for IgG Antibodies
- 06:19Amplifying, Cloning and Sequencing Heavy and Light Chain DNA
- 07:26Producing Antibodies in HEK Cells
- 08:20Anticipated Results
- 09:40Conclusion
Synthesis of human monoclonal antibodies is the first step in studies aimed at unraveling the pathophysiological mechanisms of auto-antibody-mediated immune responses. We have developed a protocol to generate recombinant human immunoglobulin G (IgG) monoclonal antibodies from blood sorted B cells, including B-cell isolation, antibody cloning and in vitro synthesis.
