Purification of Foot and mouth disease virus nonstructural protein 3ABC from vaccine in-process samples and their characterization

Foot and mouth disease is a highly contagious disease that affects the cloven hoofed animals which includes livestock and can impact on nation's economy. Adjuvanted inactivated whole virus (140s) is used as vaccine to protect animals and control the disease. The vaccine production process is well established by many commercial manufacturers by culturing the virus on BHK cell line. The development of vaccine requires partial purification of the antigen batches to eliminate unwanted proteins along with virus non structural protein 3ABC to claim as a marker vaccine in differentiating vaccinated from infected animals (DIVA) and also to improve the quality of the vaccine thus enhancing the levels of protection. In the current study, we have shown the process of eliminating 3ABC protein from the antigen batches by size exclusion chromatography using Sephacryl S-400 and Toyopearl HW 65F columns. We have demonstrated the removal of 3ABC protein in various antigen batches of FMDV Serotype O by each of the above chromatographic media and quantifying the 3ABC protein by ELISA during the chromatographic process. The process involving the treatment of sample with the buffer containing Triton X-100 has shown a minimum NSP levels in the final antigen purified by both the chromatographic media. We also suggest that the method could also be applied for other serotypes of FMDV. The present method has the potential for a large scale production of marker vaccine which can be used in differentiating vaccinated from infected animals (DIVA).