RESUMEN
The number of Hybridomes obtained from various sources rapidly increases at present. Clones producing monoclonal antibodies to influenza virus A/USSR/090/77 and to VEE-230 are generated in the laboratory of the Institute of Virology (Academy of Sciences of the USSR). The present work is devoted to the study of Hybridome karyotype by means of C-method for chromosome staining with the aim to reveal specific characteristics of these lines. Results of the investigation have shown that the count of chromosomes together with an examination of their C-staining picture permit proving a hybrid nature of the clones and identifying various Hybridomes by chromosome markers.
Asunto(s)
Anticuerpos Monoclonales/biosíntesis , Anticuerpos Antivirales/biosíntesis , Antígenos Virales/inmunología , Hibridomas/ultraestructura , Virus de la Influenza A/inmunología , Animales , Línea Celular , Bandeo Cromosómico , Virus de la Encefalitis Equina Venezolana/inmunología , Marcadores Genéticos , Hibridomas/inmunología , Inmunización , Cariotipificación , Metafase , RatonesRESUMEN
Six hybrid clones producing monoclonal antibodies (MCA) to influenza A/FMI/47 virus and 9 hybrid clones producing MCA to influenza A/USSR/090/77 virus have been developed. The resulting MCA have been shown to be highly specific for influenza viruses of H1N1 serotype and differentiating H1N1 viruses from H0N1 and H3N2. In 3 out of 11 hybrid clones under study, unique marker chromosomes not occurring in cells of the myeloma parents have been found.
Asunto(s)
Anticuerpos Monoclonales/biosíntesis , Anticuerpos Antivirales/biosíntesis , Hibridomas/inmunología , Virus de la Influenza A/inmunología , Animales , Anticuerpos Monoclonales/análisis , Anticuerpos Antivirales/análisis , Técnica del Anticuerpo Fluorescente , Pruebas de Inhibición de Hemaglutinación , Ratones , Ratones Endogámicos BALB CRESUMEN
To detect the hybrid cells forming as a result of fusion of the mouse myeloma cells with the immunic mouse splenocytes, a radioautographic method was used which involved the application of 3H-hypoxanthine as a labelled precursor of nucleic acids and of high-sensitive UK emulsion for accelerated preparations of autographs. The optimal conditions of hybridization providing for the maximum yield of hybrid cells and hybrid clones in our experiments were: use of polyethylene glycol ("Löba", mol. weight 4000) in 50% (w/v) concentration, of NS-O or X-653 myeloma cells in the ratio of parental cells 1:5 (myeloma cell: splenocyte).