| Active immunization against genetically engineering live vector vaccine of somatostatin can promote animal rapid growing. For the low virus t itre and expression in cell, the current vaccine production process is still inoculumed by chicken embryo. To improving the technological process, an established cell line named Du and Ji Rabbit kidney(DJRK) was used to culture recombinant vaccina virus. This cell line is characteristic of rapid growth, the high seeding and plating efficiencies, the dependency on serum of cell division lowers, growing in the media without serum, and so on. W/SS was inoculated onto DJRK cell culture. Specific and stable cytopathic effect was observed after 2 passages. Comparison of amplification and expression of W/SS between chicken embryo and cell indicates that the virus titre and potency are conformed to the requirements of regulations. It was obvious that the new processing technology of cell cultured is applicable to production of W/SS.The expression of fusion protein HBsAg/SS reflected the amplification of W/SS on DJRK cell line. So. A double antibody sandwich ELISA was established to measure the carrier protein HBsAg. Other, A sandwich ELISA was developed to detect SS. To gain purified special rabbit anti-SS sera IgG,3 rabbit were immunized against Trx/SS. Antibody titers were determined using indirect ELISA. Indirect inhibitor ELISA was used to detect the anti-SS specificity of the serum. The result indicated these methods are sensitive and can meet for the need of detection.Rapid measurement of antibody titre during the immunization period is essential to monitor the effectiveness of immunization protocol . The present study was designed to develop a sensitive EL ISA method to measure anti-SS and HBsAb titre values. Trx-SS was purified by osmotic shock method as coating antigen. An anti body capture ELISA system was developed to measure anti-somatostatin antibody titre. Chess titration test obtained optimal working requirement. The ELISA system was sensitive in measuring anti-somatostatin titre values. Also the HRP-SPA ELISA was established for detecting SS and HBsAg anti body of kinds of animal. HRP-SPA was used as second antibody. It showed that the ELISA had goodrepeativity, high specif ity and sensitivity. It was successfully applied for detecting antibody of SS and HBsAg of certain animals.Titrating infective virus by plaque assay is one of the most accurate methods which could determind the number of infective virus.Certain of factors,such as the volume of virus inoculum, residua of MEM, time and temperature of absorption,and so on, have remarkable influence on virus titrating. There has a unlinear relationship between the titer of virus and the volume of inoculum, the volume of liquid medium. Agar overlay has no infuence on the number of PFU before 90 h.To identify whether the passage DJRK cell line can be applied for vaccine produce. So its neop I ast i genes was detected by the nude mice. DJRK cell lines were cultured and harvested during logarithm growth, and then were counted and resuspended sterily before it was inoculated against nude mice. After a period of growth innude mice, this cell was evaluated for neop lastigenes is by pathoanotomy and pathomorphology. All of the nude mice had neoplasms after inoculation by the DJRK passaging cells. The neoplasms were tissue by pathological morphology, and their oncogene ratio was 100%. DJRK belong to oncogenous cells. It can produce malignant tumor in nude mice after inoculation subcutaneously from 20 to 40 days. |
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