[Preparation and application of mouse anti-AEG-1 monoclonal antibody]

Objective To prepare the fluorescein isothiocyanate (FITC)-labeled mouse monoclonal antibody (mAb) against astrocyte elevated gene-1 (AEG-1) and evaluate its binding ability to tumor cells. Methods The mouse anti-AEG-1 mAb was purified from mouse ascites. The purity and activity of the mAb were identified by SDS-PAGE, Western blotting and immunohistochemistry (IHC). The FITC labeling efficiency was examined by flow cytometry (FCM) and the binding ability of FITC-labeled mAb to tumor cells was detected by FCM and IHC. Results The AEG-1 mAb was successfully purified form mouse ascites, and successfully labeled by FITC. FCM showed that the fluorescence intensity of tumor cells bound with FITC- mAb was significantly strengthened, and the binding rate reached more than 90%. Moreover, the FITC-labeled anti-AEG-1 mAb could specifically recognize and bind to AEG-1 positive tumor cells. Conclusion The mouse anti-AEG-1 mAb has been successfully purified and labeled by FITC, which can specifically bind to tumor cells in vivo and in vitro.