Purification of Porcine Parvovirus by Gel Filtration

The pharmaceutical industry is required to have unit operations that remove biological contaminants, such as viruses. Porcine parvovirus (PPV) is a virus found in the majority of unvaccinated pigs around the world causing reproductive problems. It is also a good model of a small and robust non-enveloped virus that could be found in human therapeutic products. The most commonly used method for PPV purification is ultracentrifugation followed by ion-exchange chromatography. The goal of this experiment was to purify PPV from cell culture using gel filtration chromatography. Unlike the commonly used techniques, this process can be used on a large laboratory scale and can produce pure virus solution in one step. The purified PPV would be used in further research on virus removal processes. PPV supernatant was loaded onto a gel filtration column packed with Sephacryl S-300 HR and the eluate was analyzed with gel electrophoresis, radioactivity tests, and infectivity assays to determine the extent of purification of the virus. After many trials, the gel electrophoresis results finally revealed independent PPV bands, which meant that the PPV was purified.