Preparation And Application Of Monoclonal Antibody Against Ovalbumin

Objective: To prepare and identify monoclonal antibody(McAb) against ovalbumin anddevelop a method for determining ovalbumin content in influenza vaccine by ELISA.Methods: Immunize Balb/c mice with the purified ovalbumin to establish the hybridomacell strains secreting specific McAbs against ovalbumin. The specific McAbs were preparedand identified, then analyzed for specificity by Western blot, titer, steric groups, class andsubclass by indirect ELISA. Purify the McAbs by Protein A affinity chromatography. Markthe McAb with the method of NaIO4. Select the McAb with high sensitivety and goodspecificity as a candidate antibody to develop an ELISA kit. The developed kit was optimizedand analyzed for stability, sensitivity and specificity. Samples were measured by the ELISAkit and compared with Serazym Ovalbumin which was commercially available reagent.Results: Three hybridoma cell strains secreting specific McAbs against ovalbumin wereestablished, named1C2,2F7and3B2respectively. All the secreted McAbs were IgG1andshowed good specificity and titers of1：105~1：106in ascites. The relative affinity was2F7>3B2>1C2.1C2and3B2were choosed as coating antibody and2F7was choosed asenzyme labeled antibody. The developed ELISA kit reached a sensitivety of31.25ng/ml andgood linear relationship（R2=0.910）was observed between detection result and ovalbumincontent. The kit showed good stability, precision and specificity.There is no difference(p>0.05) between the results of the sample which detected by the ELISA kit and the SerazymOvalbumin reagent.Conclusion: The specific McAb against ovalbumin was successfully prepared, and anELISA kit for ovalbumin was developed, which had good sensitivity and specificity andmight be used for determining ovalbumin content in influenza vaccine.