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首页> 外文期刊>Journal of Biotechnology >Highly protective E2-CSFV vaccine candidate produced in the mammary gland of adenoviral transduced goats
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Highly protective E2-CSFV vaccine candidate produced in the mammary gland of adenoviral transduced goats

机译:在腺病毒转导山羊的乳腺中产生高度保护性的E2-CSFV候选疫苗

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摘要

Classical swine fever virus is the etiological agent of the most economically important highly contagious disease of swine worldwide. E2 is the major envelope glycoprotein present as a homodimer on the outer surface of the virus and represents an important target for the induction of neutralizing immune response against the viral infection. The E2 extracellular domain was expressed in the milk of adenoviral transduced goats at the highest level about 1.2g/L. The recombinant glycoprotein was purified from clarified serum milk by a single metal chelate affinity chromatography step, as a homodimer of approximately 100kDa and purity over 98%. Glycosylation analysis showed the presence of oligomannoside, hybrid and complex type N-glycans, attached to the recombinant E2. The capacity of goat milk-derived E2 antigen to protect pigs from both classical swine fever clinical signs and viral infection was assessed in a vaccination and challenge trial. The immunized pigs became protected after challenge with 10(5) LD(50) of a highly pathogenic CSFV strain. In the context of veterinary vaccines, this expression system has the advantages that the recombinant antigen could be harvested in about 48h after adenoviral transduction with expression levels in the range of g/L. This approach may turn into a scalable expression system for the assessment and production of veterinary vaccines.
机译:古典猪瘟病毒是全球范围内最经济重要的高度传染性猪病的病原体。 E2是主要的包膜糖蛋白,以同型二聚体的形式存在于病毒的外表面,并且是诱导针对病毒感染的中和免疫应答的重要靶标。 E2胞外域在腺病毒转导的山羊乳中以最高约1.2g / L的水平表达。通过单金属螯合亲和层析步骤从澄清的血清乳中纯化重组糖蛋白,其为约100kDa的同型二聚体且纯度超过98%。糖基化分析表明,存在连接到重组E2的寡甘露糖苷,杂合和复杂的N型聚糖。在疫苗接种和攻击试验中评估了羊奶来源的E2抗原保护猪免受经典猪瘟临床症状和病毒感染的能力。经高致病性CSFV株的10(5)LD(50)攻击后,免疫猪受到保护。在兽用疫苗的情况下,该表达系统的优点是重组病毒可以在腺病毒转导后约48小时内收获,表达水平在g / L范围内。这种方法可能会变成可扩展的表达系统,用于评估和生产兽用疫苗。

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