Efficient generation of monoclonal antibodies from single human B cells by single cell RT-PCR and expression vector cloning
T Tiller, E Meffre, S Yurasov, M Tsuiji… - Journal of …, 2008 - Elsevier
We have developed an efficient strategy that combines immunoglobulin (Ig) gene repertoire
analysis and Ig reactivity profiling at the single cell level. Based on surface marker
expression individual cells at different stages of human B cell development are isolated by
fluorescence-activated cell sorting. For each cell Ig heavy and corresponding Ig light chain
gene transcripts are amplified by nested RT-PCR and cloned into eukaryotic expression
vectors to produce monoclonal human antibodies of the same specificity in vitro. All …
analysis and Ig reactivity profiling at the single cell level. Based on surface marker
expression individual cells at different stages of human B cell development are isolated by
fluorescence-activated cell sorting. For each cell Ig heavy and corresponding Ig light chain
gene transcripts are amplified by nested RT-PCR and cloned into eukaryotic expression
vectors to produce monoclonal human antibodies of the same specificity in vitro. All …