碩士 / 國立宜蘭大學 / 動物科技學系碩士班 / 98 / Classical swine fever (CSF), also known as hog cholera, is a serious disease of pigs caused by the CSF virus (CSFV). CSF is a highly contagious disease with high mortality. Currently, vaccines for preventing classical swine fever are mainly live vaccines produced with the well-known lapinized Chinese strain (LPC) of CSFV grown in either cultured primary cells or immortalized cell lines. For many years, these vaccines have worked effectively, but due to their low market prices, production cost still needs to be lowered. Tidal bioreactor is a novel oscillating bioreactor capable of high-density cell culture for the production of recombinant proteins or viruses. In this study, several trials using a novel tidal bioreactor system to cultivate ST cell requiring only low amounts of serum for CSFV live vaccine production were performed. First, using low-serum containing medium to adapt ST cell, and test the cell growth rate and cell survival rate. Initial results for the growth of CSFV in ST cells using low-serum containing medium showed CSFV viral titers of 106TCID50/ml. Further studies showed that using the tidal bioreactor system, CSFV could be harvested for 10 batches, and with viral titers of at least 106TCID50/ml, comparable to when using medium with 5% serum. To detect any mutations during extended culturing, the sequences of highly variable region E2 gene were compared between the original isolated vaccine virus strain, the 10th batch of the tidal bioreactor harvest, and CSFV after 10 passages. The results showed that within the full-length 1120 bps E2 gene, only three nucleotides mutated, leading to only two amino acid mutations. In addition, to determine any differences in immunogenicity between these CSFV, rabbits were immunized with these three viruses and in 1, 2, 3, 4, and 5 weeks after immunization, neutralizing antibodies were measured. Although individual rabbits showed inherent differences, it was observed that at three weeks, the antibody titers of all serum increased significantly. Furthermore, all three classical swine fever virus tested produced antibodies with similar virus neutralization capacity. A platform technology was established to produce CSFV live vaccines successfully with low-serum medium and to harvest in multiple batches.
| Identifer | oai:union.ndltd.org:TW/098NIU00289008 |
| Date | January 2010 |
| Creators | Chia-yi Kao, 高嘉宜 |
| Contributors | Tsun-Yung Kuo, 郭村勇 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 77 |
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