One-step method for establishing 8-azaguanine-resistant hybridomas suitable for the preparation of triomas.
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.
Kontsekova
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E
Institute of Virology, Slovak Academy of Sciences, Bratislava, Czechoslovakia.
Novak
M
M
Macikova
I
I
Kontsek
P
P
D000818:Animals; D000918:Antibody Specificity; D001375:Azaguanine / Q000494:pharmacology*; D002459:Cell Fusion; D004351:Drug Resistance; D006735:Horseradish Peroxidase / Q000276:immunology; D006825:Hybridomas / Q000187:drug effects*; D066298:In Vitro Techniques; D051379:Mice; D011820:Rabies virus / Q000276:immunology; D013154:Spleen / Q000166:cytology
D006735:Horseradish Peroxidase; D001375:Azaguanine