Biotechnology Research Institute, National Research Council of Canada, Montreal, Canada.
Vaccine. 2012 Jan 5;30(2):300-6. doi: 10.1016/j.vaccine.2011.10.095. Epub 2011 Nov 8.
Rabies virus is an important causative agent of disease resulting in an acute infection of the nervous system and death. Although curable if treated in a timely manner, rabies remains a serious public health issue in developing countries, and the indigenous threat of rabies continues in developed countries because of wildlife reservoirs. Control of rabies in wildlife is still an important challenge for governmental authorities. There are a number of rabies vaccines commercially available for control of wildlife rabies infection. However, the vaccines currently distributed to wildlife do not effectively immunize all at-risk species, particularly skunks. A replication competent recombinant adenovirus expressing rabies glycoprotein (AdRG1.3) has shown the most promising results in laboratory trials. The adenovirus vectored vaccine is manufactured using HEK 293 cells. This study describes the successful scale-up of AdRG1.3 adenovirus production from 1 to 500 L and the manufacturing of large quantities of bulk material required for field trials to demonstrate efficacy of this new candidate vaccine. The production process was streamlined by eliminating a medium replacement step prior to infection and the culture titer was increased by over 2 fold through optimization of cell culture medium. These improvements produced a more robust and cost-effective process that facilitates industrialization and commercialization. Over 17,000 L of AdRG1.3 adenovirus cultures were manufactured to support extensive field trials. AdRG1.3 adenovirus is formulated and packaged into baits by Artemis Technologies Inc. using proprietary technology. Field trials of AdRG1.3 rabies vaccine baits have been conducted in several Canadian provinces including Ontario, Quebec and New Brunswick. The results from field trials over the period 2006-2009 demonstrated superiority of the new vaccine over other licensed vaccines in immunizing wild animals that were previously difficult to vaccinate.
狂犬病病毒是一种重要的病原体，可导致急性神经系统感染和死亡。虽然及时治疗可以治愈，但在发展中国家，狂犬病仍然是一个严重的公共卫生问题，由于野生动物宿主的存在，发达国家仍存在狂犬病本土威胁。控制野生动物中的狂犬病仍然是政府当局的一个重要挑战。有许多狂犬病疫苗可用于控制野生动物狂犬病感染。然而，目前分发给野生动物的疫苗并不能有效地使所有处于危险中的物种免疫，特别是臭鼬。表达狂犬病糖蛋白的复制型重组腺病毒（AdRG1.3）在实验室试验中显示出最有前途的结果。腺病毒载体疫苗是使用 HEK 293 细胞制造的。本研究描述了从 1 升到 500 升规模扩大 AdRG1.3 腺病毒生产的成功，并制造了大量用于现场试验的大量散装材料，以证明这种新候选疫苗的疗效。通过在感染前消除培养基替换步骤并通过优化细胞培养基将培养物滴度提高 2 倍以上，简化了生产过程。这些改进产生了更稳健且更具成本效益的工艺，有利于工业化和商业化。生产了超过 17000 升的 AdRG1.3 腺病毒培养物，以支持广泛的现场试验。AdRG1.3 腺病毒由 Artemis 技术公司使用专有的技术配制并包装成诱饵。AdRG1.3 狂犬病疫苗诱饵的现场试验已在包括安大略省、魁北克省和新不伦瑞克省在内的几个加拿大省份进行。2006 年至 2009 年期间的现场试验结果表明，与其他许可疫苗相比，新疫苗在免疫先前难以接种的野生动物方面具有优越性。
