Generation of a large complex antibody library from multiple donors

We have generated a large complex library of single chain antibodies based on four individual libraries from each of 50 donors. DNA coding for the heavy and light chain variable domains of the IgM and IgG repertoires was amplified by PCR using two different sets of primers. Each individual library w...

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Veröffentlicht in:Journal of immunological methods 1999-12, Vol.231 (1), p.3-9
Hauptverfasser: Little, Melvyn, Welschof, Martin, Braunagel, Michael, Hermes, Ingrid, Christ, Christiane, Keller, Armin, Rohrbach, Petra, Kürschner, Timo, Schmidt, Stefanie, Kleist, Christian, Terness, Peter
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Sprache:eng
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Zusammenfassung:We have generated a large complex library of single chain antibodies based on four individual libraries from each of 50 donors. DNA coding for the heavy and light chain variable domains of the IgM and IgG repertoires was amplified by PCR using two different sets of primers. Each individual library was composed of approximately 1–5×10 7 independent clones giving a final combined library of 4×10 9 members. Screening this library by phage display of single chain antibodies with small haptens, peptides and proteins yielded specific antibodies for each class of antigen.
ISSN:0022-1759
1872-7905
DOI:10.1016/S0022-1759(99)00164-7