Kontsekova E, Novak M, Macikova I, Kontsek P
Institute of Virology, Slovak Academy of Sciences, Bratislava, Czechoslovakia.
J Immunol Methods. 1991 Dec 15;145(1-2):247-50. doi: 10.1016/0022-1759(91)90333-b.
A simple and rapid one-step method for establishing azaguanine resistant (Agr) hybridomas, which can be used as a fusion partner for the construction of triomas (hybridoma x splenocyte), has been developed. The method relies on cloning the hybridoma cells in soft agar supplemented with 20 micrograms/ml 8-azaguanine. The drug-resistant subclones were isolated after 3-5 days, in comparison with 4-5 weeks reported for the conventional adaptation method. The high frequency (about 10(-3) of Agr-mutants achieved by the cloning method was demonstrated with five different hybridoma clones. One of the derived Agr-hybridomas was fused with mouse immune spleen cells in order to demonstrate its suitability for the generation of triomas secreting bispecific monoclonal antibodies.
