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sf9 昆虫细胞悬浮培养的研究
Thesis Advisor李佐虎
Degree Grantor中国科学院研究生院
Other AbstractInsect cell-baculovirus culture can be used to produce many biologicals such as biopesticide and therapeutical proteins. As a vector for foreign gene, baculovirus has the following advantages: high expression level, post-translation modification and insertion of large foreign gene. Therefore, many researchers focus their attention on this expression system which has a good commercial prospect. The following studies about the insect cell-baculovirus expression system were done: 1. In batch culture with TC-100 media (10% serum and 2.5g/l TPB), the specific growth rate was 0.023±0.003h~(-1) (95% confidence) and the specific glucose consumption rate was 13±2μg/10~6 cells·h. The cell density 2.0×10~6 cells/ml was obtained in the same media. Normally, the specific glucose consumption rate decreased with growth slowing before the maximum cell density and decreased to 0 or very low level after the maximum cell density. In normal culture, Cell growth stopped with the depletion of glucose. Lactate slightly or hardly accumulated and was consumed after the middle of culture. The abnormal growth of sf9 insect cells was usually accompanied with the abnormal lactate accumulation and the decrease of the energy metabolic efficiency. According to the characteristics of sf9 insect cell growth and glucose consumption, the phenomenological mathematical model was suggested and fitted the experiments very well. 2. In sparging culture, even if cell growth was interrupted by four hours power failure, the cell growth and glucose, lactate metabolism were not significantly different from the control. The volummetric air flow rate had the crucial effect on the cell growth. In the large volummetric air flow rate, the cell was significantly damaged and maybe changed the requirement for nutrient. At the condition of meeting the cell oxygen demand, one-order cell death rate model was suggested and the factors concerning the death rate were discussed according to this model. 3. The pro-UK yield 1065IU/ml and 1120IU/10~6 cells at cell basis in sf9 insect cell-baculovirus expression system were achieved. The pro-UK expression level increased significantly when the cell was at good state because the yield at cell basis increased significantly and there were more cells to be infected at the same nutrient depth. Therefore, the good cell state was very important to achieve the high expression level. 4. In the hydrostatic pressure oscillation of the amplitude 0.007-0.45kgf/cm~2 and cycle 4min(2min high pressure and 2min low pressure), sf9 insect cell growth and the metabolism of glucose and lactate were insignificantly different from the control. So did the Pro-UK expression.
Document Type学位论文
Recommended Citation
GB/T 7714
孙洪亮. sf9 昆虫细胞悬浮培养的研究[D]. 中国科学院研究生院,1998.
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