RESUMEN
The performance of a protein-free medium was compared in culture flasks with a serum-supplemented medium and with a serum free medium in terms of cell growth and monoclonal antibody production by a murine hybridoma. We present results of continuous production in hollow fiber culture systems using serum-free medium and protein-free medium. In protein-free medium, it has been possible to produce large quantities of monoclonal antibody with a productivity similar to that obtained in serum-free medium. After a two steps purification process, monoclonal antibodies were characterized by SDS-PAGE, High Performance Size Exclusion Chromatography and Free Solution Capillary Electrophoresis. SDS-PAGE and high performance chromatography analysis have showed that purified monoclonal antibodies produced in serum-free medium or protein-free medium were similar. Furthermore, Capillary Electrophoresis characterization revealed that both MAbs were constituted by three isoforms with equivalent electrophoretic mobilities.
Asunto(s)
Anticuerpos Monoclonales/biosíntesis , Inmunoglobulina G/biosíntesis , Animales , Anticuerpos Monoclonales/aislamiento & purificación , Recuento de Células , División Celular , Cromatografía en Gel , Medio de Cultivo Libre de Suero , Técnicas de Cultivo/instrumentación , Técnicas de Cultivo/métodos , Hibridomas , Inmunoglobulina G/aislamiento & purificación , Cinética , Ratones , Consumo de Oxígeno , Factores de TiempoRESUMEN
The human blood group Rh comprises erythrocyte membrane antigens which include the G antigen. In order to carry out this study, the G antigen was specifically immunofixed with a human monoclonal anti-Rh(G) antibody. After extraction and purification of the antigenic complex, a migration in SDS-PAGE showed the G antigen to be associated with a polypeptide of 28,000 MW.
Asunto(s)
Sistema del Grupo Sanguíneo Rh-Hr/inmunología , Anticuerpos Monoclonales/inmunología , Cromatografía de Afinidad , Electroforesis en Gel de Poliacrilamida , Eritrocitos/inmunología , Humanos , Peso MolecularRESUMEN
The G antigen is one of the erythrocyte membrane Rh antigens. The amount of Rh antigen present on the red blood cell is about 10(-15) g and radioactive labeling of membrane proteins is a useful method for its identification and characterization. In this paper, we compare 4 labeling techniques. Using a human monoclonal anti-Rh(G) antibody and an immunofixation technique, we located the G antigen on a polypeptide of an average molecular weight of 28,000 Da.
