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  • Antigen-specific monoclonal antibodies isolated from B cells expressing constitutively active STAT5.

Antigen-specific monoclonal antibodies isolated from B cells expressing constitutively active STAT5.

PloS one (2011-04-29)
Ferenc A Scheeren, Caroline M M van Geelen, Etsuko Yasuda, Hergen Spits, Tim Beaumont
ABSTRACT

Fully human monoclonal antibodies directed against specific pathogens have a high therapeutic potential, but are difficult to generate. Memory B cells were immortalized by expressing an inducible active mutant of the transcription factor Signal Transducer and Activator of Transcription 5 (STAT5). Active STAT5 inhibits the differentiation of B cells while increasing their replicative life span. We obtained cloned B cell lines, which produced antibodies in the presence of interleukin 21 after turning off STAT5. We used this method to obtain monoclonal antibodies against the model antigen tetanus toxin. Here we describe a novel and relatively simple method of immortalizing antigen-specific human B cells for isolation of human monoclonal antibodies. These results show that STAT5 overexpression can be employed to isolate antigen specific antibodies from human memory B cells.

MATERIALS
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Sigma-Aldrich
Interleukin-21 from mouse, ≥97% (SDS-PAGE), recombinant, expressed in E. coli, lyophilized powder, suitable for cell culture
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