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PLoS One. 2011 Apr 15;6(4):e17189. doi: 10.1371/journal.pone.0017189.

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

Author information

1
Department of Cell Biology and Histology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands.

Abstract

BACKGROUND:

Fully human monoclonal antibodies directed against specific pathogens have a high therapeutic potential, but are difficult to generate.

METHODOLOGY/PRINCIPAL FINDINGS:

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.

CONCLUSIONS/SIGNIFICANCE:

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.

PMID:
21525985
PMCID:
PMC3078118
DOI:
10.1371/journal.pone.0017189
[Indexed for MEDLINE]
Free PMC Article

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