A new and rapid method for the selection and cloning of antigen-specific hybridomas with magnetic microspheres.

Journal of Immunological Methods
J K HortonS Swinburne

Abstract

A new and highly selective procedure is described for the rapid selection and cloning of antibody-secreting hybridomas with antigen-coated magnetic beads. Immune splenocytes were fused to Sp2 myeloma cells with a PEG/DMSO mixture. Cells were cultured in HAT and screened for the presence of specific antibody after 7-10 days. Hybridomas from the positive colonies were mixed with antigen-coated magnetic polymer beads and antigen-specific cells separated on a magnet. Cloning was carried out directly by limiting dilution with the magnetic beads still bound on the cells. No obvious toxic effects were observed. The antibodies established by this technique were of a high affinity (greater than 10(9) l/mol) and were generated in 50% of the usual process time. The procedure described here should greatly facilitate the process of obtaining hybridomas and expand the range of monoclonal antibodies available.

References

Apr 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·D R ParksL A Herzenberg

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Citations

Dec 31, 1990·Journal of Immunological Methods·J K HortonM J O'Sullivan
Jul 6, 1993·Journal of Immunological Methods·P G SteenbakkersW Olijve
Apr 1, 1991·Current Opinion in Immunology·F L Battye, K Shortman
Jun 6, 1997·Journal of Neuroscience Methods·A P WrightR J Colello
Sep 1, 1993·Scandinavian Journal of Immunology·A Bennick, F Brosstad
Jun 3, 2010·Biotechnology Journal·Prabhjyot Kaur DehalDavid John Pritchard

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