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Flow cytometry histogram showing human mononuclear cells incubated with anti-CD4 biotin and followed by staining with PE-Cy7 streptavidin.
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Abstract
Rapid generation of fully human monoclonal antibodies specific to a vaccinating antigen.
Authors:Smith K, Garman L, Wrammert J, Zheng NY, Capra JD, Ahmed R, Wilson PC,
Journal:Nat Protoc
PubMed ID:19247287
We describe herein a protocol for the production of antigen-specific human monoclonal antibodies (hmAbs). Antibody-secreting cells (ASCs) are isolated from whole blood collected 7 d after vaccination and sorted by flow cytometry into single cell plates. The antibody genes of the ASCs are then amplified by RT-PCR and nested PCR, ... More