ABSTRACT
Objective To prepare a neutralizing monoclonal antibody against rabies virus. Meth-ods BALB/c mice were immunized with the inactivated rabies virus CTN strains on day 0, 7, 14 and 28. Spleen cell samples were collected and then fused with SP2/0 cells to prepare the hybridoma cell line. Posi-tive hybridoma cells that were screened out with RFFIT technique were injected into BALB/c mice intraper-itoneally. Ascites samples were collected from the mice to separate neutralizing monoclonal antibodies. Affin-ity chromatography was used for the purification of neutralizing monoclonal antibodies. Subtype identification and sequencing analysis were performed for further identification. A colloidal gold strip based method for rap-id detection of rabies vaccine was established with the prepared monoclonal antibodies. Results The hybri-doma cell line, CTN-McAb1, was prepared successfully with stable secretion of neutralizing monoclonal anti-bodies against rabies virus. The purity of those antibodies was more than 95% after purification and the sub-type of them was IgG1. The colloidal gold strip for raid detection of rabies vaccine was successful prepared. Conclusion The neutralizing monoclonal antibody against rabies virus was successfully prepared and could be used for preliminary application. This study will be of great significance for the quality control of rabies vaccine.
ABSTRACT
Objective To explore the application of atomic force microscopy( AFM ) on the research of morphology of the rabies viruses. Methods To prepare the rabies virus CTN-1v strains by ultracentrifugation, and observe it with transmission electron microscopy (TEM) which negatively stained by phosphotungstic acid. Then study the morphology of rabies virus with AFM based on the result of TEM. AFM image applies the tapping mode to rabies virus without any further treatment in air at room temperature. Results The TEM image is two-dimensional image which can be seen the classical bullet-shaped structure,and the spike structure can also be seen. The AFM image showed the rabies virus morphology with three-dimensional image which can shows the characteristics of the virus surface and edge. The rabies virus particle was successfully observed by TEM or AFM methods. Conclusion It's the first time to get the three-dimensional morphological structure of rabies virus by atomic force microscopy, compared with transmission electron microscopy, AFM is a new research tool for viral morphology study with the advantages of simple sample preparing and intuitionistic and visible interface for researchers.