Development of a method for detection of neutralizing activity of antibody against novel recombinant human erythropoietin

Objective To develop a method for detection of neutralizing activity of antibody against novel recombinant human erythropoietin(rHuEPO)and apply to analysis of samples in toxicological test.Methods The promoting effect of novel rHuEPO on proliferation of human UT7/EPO cells as well as the inhibitory effect of antibody(rabbit antiserum)against the novel rHuEPO on the proliferation were evaluated,based on which a method for detection of neutralizing activity of antibody against novel rHuEPO was developed and used for relevant antibodies in toxicological test.Results The novel rHuEPO at a concentration of 0.125 ～ 128 ng/ml showed significantly promoting effect on the proliferation of UT7/EPO cells.However,the antibody against novel rHuEPO,at a concentration of 10-1 ～ 10-4,showed significantly inhibitory effect on the proliferation,and the inhibition curve was of characteristics of neutralizing antibody.In toxicological test,SD rat serum against novel rHuEPO showed significantly specific inhibitory effect,while Beagle canine serum against novel rHuEPO showed no significant effect,on the proliferation of human UT7/EPO cells.Conclusion A method for detection of neutralizing activity against novel rHuEPO was successfully developed and used for analysis of samples in toxicological test,which provided an effective tool for evaluation of immunogenicity of novel rHuEPO in long-term toxicity test.