One-step method for establishing 8-azaguanine-resistant hybridomas suitable for the preparation of triomas
Language English Country Netherlands Media print
Document type Journal Article
PubMed
1765658
DOI
10.1016/0022-1759(91)90333-b
PII: 0022-1759(91)90333-B
Knihovny.cz E-resources
- MeSH
- Azaguanine pharmacology MeSH
- Cell Fusion MeSH
- Hybridomas drug effects MeSH
- Horseradish Peroxidase immunology MeSH
- Drug Resistance MeSH
- Mice MeSH
- Spleen cytology MeSH
- Antibody Specificity MeSH
- In Vitro Techniques MeSH
- Rabies virus immunology MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Azaguanine MeSH
- Horseradish Peroxidase MeSH
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.
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