Stabilisation of Newcastle Disease vaccine formulated in sugar-glass on polypropylene membranes

聚丙烯膜上糖玻璃配制的新城疫疫苗的稳定性

• 批准号: BB/M019152/1
• 负责人: Adrian Hill
• 金额: $ 17.95万
• 依托单位: University of Oxford
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2015
• 资助国家: 英国
• 起止时间: 2015 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FM019152%2F1
• 关键词: Stabilisation; Newcastle; Disease; vaccine; formulated

Newcastle disease (ND) is a highly contagious disease affecting poultry, with diverse symptoms involving the nervous, respiratory and digestive systems. Mortality rates depend on the particular viral strain and can vary from 0 to 100%; Newcastle Disease Virus (NDV) occasionally infects humans with mild symptoms e.g. conjunctivitis. The disease presents a major issue for poultry farmers, and is endemic in many countries of the world. Vaccines have been developed against NDV, usually based on non-virulent (weakened) forms of the virus. The vaccine is administered in drinking water or by aerosol spray to enable large flocks to be protected simultaneously, or can be dosed by eye drops. NDV is a particularly unstable pathogen because its genetic material is made from RNA, rather than DNA - this means that the vaccine needs to be stored by refrigeration in a lyophilised (dry powder) state. As a result, many farmers with small flocks of birds, typically chickens, in low-resource countries of the world do not have access to vaccine and can lose entire flocks on a regular basis. The problem is further compounded by the fact that the vaccine is manufactured in vials containing 500-1000 doses, and is therefore too expensive for use in subsistence farming. Lack of thermo-stability is a serious problem for many vaccines, both human and animal, as developing countries with no reliable 'cold chain' (network of refrigerated storage) may not be able to deliver active vaccines to remote regions and vaccine wastage is considerable.This project is a joint venture between the University of Oxford and the Pirbright Institute, both experienced in the development of new vaccines. Our objective is to develop a technology for thermo-stabilisation of NDV vaccine, which in the longer term can be applied to other vaccines. Lyophilisation (drying to form a powder) is currently used to stabilise many vaccines, but is not ideal as i) the manufacturing process results in substantial losses and includes freezing and drying steps which can adversely affect viability, and ii) all lyophilised licensed vaccines require refrigeration. We plan to use a stabilisation technology called 'sugar-glass' formulation which involves mixing vaccines with a concentrated solution of sugar, and then drying onto fibrous membranes to form inert sugar-glass which thinly coats the membrane fibres. Vaccines are quickly and simply resuspended after storage by the addition of buffer to the membrane. The technology has been tested previously with an experimental malarial vaccine but further development was impeded by lack of a suitable support matrix.We will aim to solve this problem by modifying Polypropylene (PP) membrane using Plasma treatment. PP is in many ways an ideal matrix for NDV vaccine sugar-glass, but is not 'wettable' and will not absorb vaccine without pre-treatment. Plasma treatment involves passing radio waves through a gas in order to produce charged ions; this 'plasma' is then used to treat surfaces including water-repellent polymers to make them wettable. The method is used in the printing industry to treat PP membrane so that it adheres to ink, but has not been applied to absorption of biological molecules. The technology is ideal for quick, cost-effective manufacture of stabilised NDV vaccine and is suitable for transfer to manufacturers in developing countries, where PP membrane can be cut into sizes appropriate for vaccination of local flocks. During the 18 month project we will a) test whether we can form a sugar-glass containing NDV vaccine on plasma treated PP membranes, b) incubate the stabilised vaccine at different temperatures and at various times, mimicking transport and storage conditions in tropical countries (4oC to 55oC), and c) test recovery of the vaccine using a variety of biological assays including the ability to induce a protective level of antibodies against the disease in chickens.

新城疫（ND）是一种影响家禽的高度传染性疾病，具有涉及神经系统、呼吸系统和消化系统的多种症状。死亡率取决于特定的病毒株，从0到100%不等；新城疫病毒（NDV）偶尔感染人类，症状轻微，例如结膜炎。该病对家禽养殖户来说是一个重大问题，在世界许多国家流行。针对新城疫的疫苗已经开发出来，通常是基于该病毒的无毒（弱化）形式。该疫苗可在饮用水中接种，或通过气溶胶喷雾接种，以使大群猪群同时得到保护，也可通过滴眼液给药。NDV是一种特别不稳定的病原体，因为它的遗传物质是由RNA而不是DNA构成的——这意味着疫苗需要以冻干（干粉）状态冷藏储存。因此，在世界上资源匮乏的国家，许多拥有小群禽鸟（通常是鸡）的农民无法获得疫苗，并可能经常失去整群禽鸟。使问题进一步复杂化的是，疫苗是装在每瓶500至1000剂的小瓶中生产的，因此对于自给农业来说过于昂贵。缺乏热稳定性是许多疫苗（包括人用疫苗和动物疫苗）面临的一个严重问题，因为没有可靠的“冷链”（冷藏储存网络）的发展中国家可能无法向偏远地区运送活疫苗，而且疫苗浪费相当大。这个项目是牛津大学和皮尔布赖特研究所之间的一个合资企业，两者都在开发新疫苗方面经验丰富。我们的目标是开发一种新冠病毒疫苗的热稳定技术，从长远来看，这种技术可以应用于其他疫苗。冻干（干燥形成粉末）目前用于稳定许多疫苗，但并不理想，因为i)生产过程会导致大量损失，包括可能对活力产生不利影响的冷冻和干燥步骤，ii)所有冻干许可疫苗都需要冷藏。我们计划使用一种称为“糖玻璃”配方的稳定技术，将疫苗与浓缩的糖溶液混合，然后干燥到纤维膜上，形成惰性的糖玻璃，薄薄的覆盖在膜纤维上。通过在膜上添加缓冲液，疫苗在储存后可以快速而简单地重悬。这项技术以前已经用一种实验性疟疾疫苗进行了测试，但由于缺乏合适的支持基质，进一步的开发受到了阻碍。我们将通过等离子体处理对聚丙烯（PP）膜进行改性来解决这一问题。PP在许多方面是NDV疫苗糖玻璃的理想基质，但它不“可湿性”，不经预处理就不能吸收疫苗。等离子体处理包括使无线电波穿过气体以产生带电离子；然后，这种“等离子体”被用来处理表面，包括防水聚合物，使其可湿性。该方法在印刷工业中用于处理PP膜，使其粘附在油墨上，但尚未应用于吸收生物分子。该技术是快速、经济高效地生产稳定的新城疫疫苗的理想技术，适合转移给发展中国家的制造商，在那里PP膜可以切割成适合当地鸡群接种的尺寸。在这个为期18个月的项目中，我们将a)测试我们是否可以在血浆处理过的PP膜上形成含有NDV疫苗的糖玻璃，b)在不同温度和不同时间孵育稳定的疫苗，模拟热带国家的运输和储存条件（4℃至55℃），以及c)使用各种生物测定法测试疫苗的恢复情况，包括在鸡体内诱导保护性抗体水平的能力。

项目成果

Characterisation of factors contributing to the performance of nonwoven fibrous matrices as substrates for adenovirus vectored vaccine stabilisation.

• DOI: 10.1038/s41598-021-00065-4
• 发表时间: 2021-10-22
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Dulal P;Gharaei R;Berg A;Walters AA;Hawkins N;Claridge TDW;Kowal K;Neill S;Ritchie AJ;Ashfield R;Hill AVS;Tronci G;Russell SJ;Douglas AD
• 通讯作者: Douglas AD