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
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.