Preparation Of Monoclonal Antibodies Against Adenoviral Penton And Development Of Sandwich ELISA

The Intestinal adenovirus can cause infantile diarrhea,it has become the second pathogen after rotavirus,so the key to found against the virus rapid and sensitive detection method to prevent and control this disease,it prepared by the anti-adenovirus penton McAb in this research,then analysis and identify its biological featurs,then it was established a double-antibody sandwich ELISA,it provides an effective method for the virus detection and prevention.The specific reserch as follows:1. Preparation and identification of Monoclonal Antibodies against adenoviral pentonUsing pronucleus genetically engineered antigen by adenoviral penton to immunize BALB/c mice,after immunized the third time,using mouse spleen lymphocytes and mouse SP2/0cells to fuse after immunization,then using indirect ELISA to screen by adenovirus penton protein pronucleus engineered antigens.It got five hybridomas after using indirect ELISA to screen hybridoma by fusing used adenovirus penton antigen,the subcategory Ig was IgG2a and IgG1,given five hybridomas serially passaged experiments in vitro for3months and serially passaged mouse ascites,they can secreted McAb sustaining,cell titers remained around2000and ascites titers remained200000stabled,it has no cross-reaction with Salmonella typhimurium and rotavirus which can caused diarrhea,with stability、specificity and high titer.2.Development of adenovirus penton Sandwich ELISA for detectionUsing horseradish peroxidase (HRP) labeled4F2McAb after purifing,with McAb4D7as capture antibody to establish for the detection of sandwich ELISA.The detection is specific for adenovirus,it also has no cross-reaction with Salmonella typhimurium and rotavirus,this research is expected to detect adenovirus in clinical.