Objective: We conducted a preliminary study on the feasibility of preparing monoclonal antibody by umbilical cord blood cells systems. Methods: Collected umbilical cord blood cells and stimulated by Rubella virus, the...Objective: We conducted a preliminary study on the feasibility of preparing monoclonal antibody by umbilical cord blood cells systems. Methods: Collected umbilical cord blood cells and stimulated by Rubella virus, then incubated them and collected cell supernatant. By using Caprylic acid- saturated Ammonium sulfate method, monoclonal antibody was purified, and IgG subtype identification was conducted by their subtype classification kit of sigma. In traditional way, the preparation of monoclonal antibody cannot do without BABL/C mice and SP2/0 hybridoma cells. Consider this kinds of monoclonal antibody as a positive control, the reactivity and specificity of monoclonal antibodies were identified by Dot-ELISA. Results: By ELISA, we obtained four strains of positive umbilical cord blood cells. After subculture, cryopreservation and resuscitation in vitro, three of them were confirmed to secrete monoclonal antibody against rubella virus stably. The result of monoclonal antibody subtype classification showed that, both IgG1 and IgG2 types were detected. However, the quantity of monoclonal antibody prepared by umbilical cord blood cells was less than that from traditional method. Conclusions: The method of prepare monoclonal antibody using umbilical cord blood cells is feasible.展开更多
文摘Objective: We conducted a preliminary study on the feasibility of preparing monoclonal antibody by umbilical cord blood cells systems. Methods: Collected umbilical cord blood cells and stimulated by Rubella virus, then incubated them and collected cell supernatant. By using Caprylic acid- saturated Ammonium sulfate method, monoclonal antibody was purified, and IgG subtype identification was conducted by their subtype classification kit of sigma. In traditional way, the preparation of monoclonal antibody cannot do without BABL/C mice and SP2/0 hybridoma cells. Consider this kinds of monoclonal antibody as a positive control, the reactivity and specificity of monoclonal antibodies were identified by Dot-ELISA. Results: By ELISA, we obtained four strains of positive umbilical cord blood cells. After subculture, cryopreservation and resuscitation in vitro, three of them were confirmed to secrete monoclonal antibody against rubella virus stably. The result of monoclonal antibody subtype classification showed that, both IgG1 and IgG2 types were detected. However, the quantity of monoclonal antibody prepared by umbilical cord blood cells was less than that from traditional method. Conclusions: The method of prepare monoclonal antibody using umbilical cord blood cells is feasible.
