Authorizing of Immunogenicity of concentrated and purified Newcastle disease virus using downstream processing

Newcastle disease virus from the Paramyxoviridae family is a single-stranded negative-sense RNA virus. NDV is an infection of domestic poultry and nearly all bird species. It has been a very severe difficulty for the poultry industry all over the world Even though it remains a potential threat to poultry industries, this virus is a powerful oncolytic virus as well. In this study a process was accomplished to achieve concentrated and highly purified NDV V۴ strain particles. Downstream processing of Newcastle virus strain V۴ was characterized by amplifying virus in embryonated chicken eggs. Through a sequence of steps, harvesting allantoic fluid, clarification by centrifuge, concentration by ultrafiltration, size exclusion separation, the reduced volume and pure virus particles were considered for the amount of ovalbumin, hemagglutinin activity, electron microscopy (TEM), electrophoresis, and additionally immunogenicity of prepared antigens. The results presented a high recovery of HA activity in concentrated and pure virus with removing of ovalbumin and the typical morphology based on TEM. Sepharose CL-۴B was determined as the best media among all used resins to purify the virus. Prepared formulations as vaccines demonstrated a positive hemagglutinin inhibition for ۶ months duration and stable for two years. Organized study provided strong evidence that this method was quite appropriate in concentrating and purifying of intact Newcastle disease virus to use in vaccine research, also in antiserum preparation, or probably like an oncotic agent as an alternative to conventional procedures. Though additional studies are being tested, this procedure can be used practically on a semi-industrial scale in the production of multiple vaccine components.

Keywords:

Newcastle virus , vaccine , Ultrafiltration , chromatography